LOCUS       SEGA062                4413 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12898.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4413)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PT7-low-TEV-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4413
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     CDS             1..528
                     /codon_start=3
                     /label=glmS
                     /translation="VHGLQALPSRIEQMLSQDKRIEALAEDFSDKHHALFLGRGDQYPI
                     ALEGALKLKEISYIHAEAYAAGELKHGPLALIDADMPVIVVAPNNELLEKLKSNIEEVR
                     ARGGQLYVFADQDAGFVSSDNMHIIEMPHVEEVIAPIFYTVPLQLLAYHVALIKGTDVD
                     QPRNLAKSVTVE"
     misc_feature    538..594
                     /label=L3S2P11 Terminator
     CDS             complement(604..1686)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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(1687..1764)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1795..1813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1814..1838
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     RBS             1850..1861
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    1862..1876
                     /label=TIR
     CDS             1868..1888
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="MENLYFQ"
     CDS             1889..2599
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006) ?"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    2603..2651
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2662..2690
                     /label=Promoter 3BCD2
     CDS             2730..3920
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    3901..3950
                     /label=HR3
     terminator      3931..3971
                     /label=rrnC
     misc_feature    3972..4021
                     /label=homology
     CDS             4166..4413
                     /codon_start=1
                     /label=pstS
                     /translation="MKVMRTTVATVVAATLSMSAFSVFAEASLTGAGATFPAPVYAKWA
                     DTYQKETGNKVNYQGIGSSGGVKQIIANTVDFGASDA"
ORIGIN
        1 gtgcatggtc tgcaggcgct gccgagccgt attgagcaga tgctgtctca ggacaaacgc
       61 attgaagcgc tggcagaaga tttctctgac aaacatcacg cgctgttcct gggccgtggc
      121 gatcagtacc caatcgcgct ggaaggcgca ttgaagttga aagagatctc ttacattcac
      181 gctgaagcct acgctgctgg cgaactgaaa cacggtccgc tggcgctaat tgatgccgat
      241 atgccggtta ttgttgttgc accgaacaac gaattgctgg aaaaactgaa atccaacatt
      301 gaagaagttc gcgcgcgtgg cggtcagttg tatgtcttcg ccgatcagga tgcgggtttt
      361 gtaagtagcg ataacatgca catcatcgag atgccgcatg tggaagaggt gattgcaccg
      421 atcttctaca ccgttccgct gcagctgctg gcttaccatg tcgcgctgat caaaggcacc
      481 gacgttgacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatctc
      541 ggtaccaaat tccagaaaag agacgctttc gagcgtcttt tttcgttttg gtccgcgttg
      601 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      661 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
      721 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
      781 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
      841 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
      901 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
      961 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     1021 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     1081 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     1141 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     1201 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     1261 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     1321 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     1381 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     1441 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     1501 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     1561 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     1621 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     1681 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     1741 ggttttgcgc cattcgatgg tgtcggatcg agatcgatct cgatcccgcg aaattaatac
     1801 gactcactat aggggaattg tgagcggata acaattccta ctagagatta aagaggagaa
     1861 acgggggatg gagaacctgt acttccagag caaaggagaa gaacttttca ctggagttgt
     1921 gccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     1981 gggtgaaggt gatgctacat acggaaaact cacccttaaa tttatttgca ctactggaaa
     2041 actacctgtt ccatggccaa cacttgtcac tactctgacc tatggcgttc aatgcttttc
     2101 ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     2161 tgtacaggaa cgcactatat ctttcaaaga tgacgggaac tacaagacgc gtgctgaagt
     2221 caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga
     2281 agatggaaac attctcggac acaaactaga gtacaactat aactcacaca atgtatacat
     2341 cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacattga
     2401 agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     2461 tgtcctttta ccagacaacc attacctgtc gacacaatct gccctttcga aagatcccaa
     2521 cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg
     2581 catggatgag ctctacaaat gaccaattat tgaaggccgc taacgcggcc tttttttgtt
     2641 tctggtctcc caaaaattta tttgcttatt aatcatccgg ctcgtataat gtgtggaggg
     2701 cccaagttca cttaaaaagg agaatagtta tgaaatcgaa caatgcgctc atcgtcatcc
     2761 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
     2821 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc
     2881 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg
     2941 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
     3001 cgaccacacc cgtcctgtgg attctctacg ccggacgcat cgtggccggc atcaccggcg
     3061 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     3121 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     3181 ggggactgtt gggcgccatc tccctgcacg caccattcct tgcggcggcg gtgctcaacg
     3241 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgccgtc
     3301 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacca
     3361 ttgtggccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     3421 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg agcggcctgt
     3481 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
     3541 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct
     3601 acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
     3661 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
     3721 accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg
     3781 gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat
     3841 ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga
     3901 gccgggccac ctcgacctga gacaattgtc cagataaaaa aaatccttag ctttcgctaa
     3961 ggatgatttc taaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat
     4021 atcaacgctt atttaaatca gactgaagac tttatctctc tgtcataaaa ctgtcatatt
     4081 ccttacatat aactgtcacc tgtttgtcct attttgcttc tcgtagccaa caaacaatgc
     4141 tttatgaatc ctcccaggag acattatgaa agttatgcgt accaccgtcg caactgttgt
     4201 cgccgcgacc ttatcgatga gtgctttctc tgtgtttgca gaagcaagcc tgacaggtgc
     4261 aggtgcaacc ttccctgcgc cggtgtatgc caaatgggct gacacttacc agaaagaaac
     4321 cggtaataaa gttaactacc agggtatcgg ttcttccggt ggcgtaaaac agattatcgc
     4381 taataccgtt gattttggtg cctctgacgc gcc
//
