LOCUS       pET11a-Chloro-ACL                8712 bp ds-DNA     circular SYN 18-SEP-2017
DEFINITION  LMBP 11125.
ACCESSION   .
VERSION     .
KEYWORDS    316_pET11a-Chloro-ACL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct

FEATURES             Location/Qualifiers
     source          1..8712
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        104..208
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             209..1069
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3761..4843)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4844..4921)
                     /gene="lacI"
                     /note="lacI promoter"
     promoter        5230..5248
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5304..5309
                     /note="ribosome binding site"
     CDS             5318..6514
                     /codon_start=1
                     /note="ACL_A"
                     /translation="MAKILEGPAMKLFNKWGIPVPNYVVIMDPDRLAQLGEANKWLRES
                     KLVVKAHEAIGGRFKLGLVKIGLNLEQAIEAAKEMLGAKVGTAEIGQVIIAEMLEHDAE
                     FYVSIIGNKDGAELLISKHGGVDIEDNWDSVRRIQIELDENPTIEQLTELAKDAGFEGE
                     IAERVGKICSRLILCFDNEDAQSIEINPLVIRKSDMRFAALDAVMNVDYDARFRHADWD
                     FKPVSEIGRPFTEAEQQIMEIDSRIKGSVKFVEVPGGEIALLTAGGGASVFYADAVVAR
                     GGTIANYAEYSGDPADWAVEALTETICRLPNIKHIIVGGAIANFTDVKATFSGIINGFR
                     ESKSKGYLEGVKIWVRRGGPNEAQGLAAIKQLQEEGFDIHVYDRSMPMTDIVDLAMKS"
     CDS             5318
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /note="T7 tag"
                     /translation=""
     CDS             6535..8388
                     /codon_start=1
                     /note="ACL_B"
                     /translation="MSILANKDTRAVIIGGVAGVNAAKRMAQFDFLVNRPLTVQAFVYP
                     PEAGQQKEIFRGGELKNVTVYDSLAPALEEHPDINTALIYLGASRAAQAAKEALESPNI
                     QLVSMITEGVPEKDAKRLKKLAQKLGKMLNGPSSIGIMSAGECRLGVIGGEFKNLKLCN
                     LYRQGSFGVLTKSGGLSNEAMWLCAQNGDGITSAVAIGGDAYPGTDFVTYLEMFEKDPA
                     TKAVVMIGEVGGNLEEEAAEWLAAEPRRIKLIAAIGGTCQEVLPQGMKFGHAGAKEGKK
                     GAGSARSKMNALRDAGAYVPDTFGGLSKEIKKVYEELIAAGEISTEIDEAVLPELPPRV
                     QEVMKQGEVIVEPLIRTTISDDRGEEPRYAGYAASELCSKGYGIEDVIGLLWNKKLPTR
                     EESEIIKRIVMISADHGPAVSGAFGSILAACAGIDMPQAVSAGMTMIGPRFGGAVTNAG
                     KYFKMAVEDYPNDIPGFLSWMKKNVGPVPGIGHRVKSVKNPDQRVKYLVSYIKNETSLH
                     TPCLDYALEVEKVTTAKKGNLILNVDGTIGCILMDLDFPVHSLNGFFVLARTIGMIGHW
                     IDQNNQNSRLIRLYDYLINYAVKPEQEVPEKKGGSHHHHHH"
     CDS             8368..8385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      8453..8500
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3721 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3841 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3901 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3961 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4021 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     4081 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4141 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4201 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4261 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4321 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4381 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4441 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4501 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4561 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4621 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4681 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4741 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4801 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4861 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4921 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     4981 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     5221 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     5281 aaataatttt gtttaacttt aagaaggaga tatacatatg gccaaaattc tggaaggtcc
     5341 ggcaatgaaa ctgtttaaca aatggggtat tccggtgccg aattatgttg ttattatgga
     5401 cccggatcgt ctggcacagc tgggtgaagc aaataaatgg ctgcgtgaaa gcaaactggt
     5461 tgttaaagca catgaagcaa ttggcggtcg ttttaaactg ggtctggtta aaattggtct
     5521 gaatctggaa caggcaattg aagcagcaaa agaaatgctg ggtgcaaaag ttggcaccgc
     5581 agaaattggt caggttatta ttgcagagat gctggaacat gatgccgagt tttatgttag
     5641 cattatcggc aataaagatg gtgccgaact gctgattagc aaacatggtg gtgttgatat
     5701 tgaggataat tgggatagcg ttcgtcgcat tcagattgaa ctggatgaaa atccgaccat
     5761 tgaacagctg accgaactgg caaaagatgc aggttttgaa ggtgaaattg cagaacgtgt
     5821 tggtaaaatt tgtagccgtc tgattctgtg ctttgataat gaagatgcac agagcattga
     5881 aatcaatccg ctggttattc gtaaaagcga tatgcgtttt gcagcactgg atgcagttat
     5941 gaatgttgat tatgatgccc gttttcgtca tgccgattgg gattttaaac cggttagcga
     6001 aattggccgt ccgtttaccg aagcagaaca gcagattatg gaaattgata gccgtattaa
     6061 aggcagcgtg aaatttgttg aagttccggg tggcgaaatt gcactgctga ccgcaggcgg
     6121 tggtgcaagc gttttttatg cagatgcagt tgttgcacgt ggtggcacca ttgccaatta
     6181 tgcagaatat agcggtgatc cggcagattg ggcagttgaa gccctgaccg aaaccatttg
     6241 tcgtctgccg aacattaaac atattattgt tggtggtgcg atcgccaatt ttaccgatgt
     6301 taaagccacc tttagcggca ttattaacgg ttttcgtgag agcaaaagca aaggttatct
     6361 ggaaggcgtg aaaatttggg ttcgtcgtgg tggtccgaat gaagcacagg gtctggcagc
     6421 aattaaacag ctgcaagaag agggttttga tatccatgtt tatgatcgta gcatgccgat
     6481 gaccgatatt gttgatctgg ccatgaaaag ctaagcattc gaaggagata tcacatgagc
     6541 atcctggcca ataaagatac ccgtgcagtt attattggtg gtgttgccgg tgttaatgca
     6601 gcaaaacgta tggcacagtt tgattttctg gttaatcgtc cgctgaccgt tcaggcattt
     6661 gtttatccgc ctgaagcagg tcagcagaaa gaaatttttc gtggtggtga actgaaaaac
     6721 gtgaccgttt atgatagcct ggcaccggca ctggaagaac atccggatat caataccgca
     6781 ctgatttatc tgggtgcaag ccgtgcagca caggcagcaa aagaagcact ggaaagcccg
     6841 aatattcagc tggttagcat gattaccgaa ggtgttccgg aaaaagatgc aaaacgtctg
     6901 aaaaaactgg cacagaaact gggcaaaatg ctgaatggtc cgagcagcat tggtattatg
     6961 tcagccggtg aatgtcgtct gggtgttatt ggcggtgagt ttaaaaacct gaaactgtgc
     7021 aatctgtatc gccagggtag ctttggtgtt ctgaccaaaa gcggtggtct gagcaatgaa
     7081 gcaatgtggc tgtgtgcaca gaatggtgat ggtattacca gcgcagttgc aatcggtggt
     7141 gatgcatatc cgggtacaga ttttgttacc tatctggaaa tgttcgagaa agatccggca
     7201 accaaagcag ttgttatgat tggtgaagtt ggtggcaatc tggaagaaga ggcagcagaa
     7261 tggctggcag ccgaaccgcg tcgtattaaa ctgattgcag caattggtgg cacctgtcaa
     7321 gaagttctgc cgcagggtat gaaatttggt catgccggtg cgaaagaagg taaaaaaggc
     7381 gcaggtagcg cacgtagcaa aatgaatgcc ctgcgtgatg cgggtgccta tgttccggat
     7441 acctttggtg gtctgtcaaa agaaatcaaa aaagtgtacg aggaactgat cgcagcgggt
     7501 gaaattagca ccgaaattga tgaagccgtt ctgccggaac tgcctccgcg tgttcaagag
     7561 gttatgaaac agggtgaagt tattgttgaa ccgctgattc gtaccaccat tagtgatgat
     7621 cgtggtgaag aaccgcgtta tgcaggttat gccgcaagcg aactgtgtag caaaggttat
     7681 ggtattgaag atgttattgg tctgctgtgg aacaaaaaac tgccgacccg tgaagaaagc
     7741 gaaattatca aacgtattgt gatgatcagt gcagatcatg gtccggcagt tagcggtgca
     7801 tttggtagca ttctggcagc atgcgcaggc attgatatgc cgcaggccgt tagcgcaggt
     7861 atgaccatga ttggtccgcg ttttggtggt gcagttacca atgcaggcaa atatttcaaa
     7921 atggccgttg aggattatcc gaatgatatt ccgggttttc tgagctggat gaaaaaaaac
     7981 gttggtccgg ttccgggtat tggtcatcgt gttaaaagcg ttaaaaatcc ggatcagcgt
     8041 gtgaaatatc tggtgagcta tatcaaaaat gaaaccagcc tgcatacccc gtgtctggat
     8101 tatgccctgg aagttgaaaa agttaccacc gccaaaaaag gcaatctgat tctgaatgtt
     8161 gatggcacca ttggctgtat tctgatggat ctggattttc cggttcatag cctgaatggt
     8221 ttttttgtgc tggcacgtac cattggtatg attggccatt ggattgatca gaataatcag
     8281 aacagccgtc tgattcgcct gtatgattat ctgattaact atgccgttaa accggaacaa
     8341 gaagtgccgg aaaaaaaagg tggtagccat catcatcacc atcactaagg atccggctgc
     8401 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     8461 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     8521 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
     8581 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
     8641 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
     8701 caaacatgag aa
//

