LOCUS       SEGA065                4449 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12901.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4449)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PT7-low-PROTi-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4449
                     /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"
     misc_feature    1889..1924
                     /label=PROTi-Linker
     CDS             1925..2635
                     /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    2639..2687
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2698..2726
                     /label=Promoter 3BCD2
     CDS             2766..3956
                     /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    3937..3986
                     /label=HR3
     terminator      3967..4007
                     /label=rrnC
     misc_feature    4008..4057
                     /label=homology
     CDS             4202..4449
                     /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 acttccagtt tcgtagcaaa ggagaagaac ttgtcactgg
     1921 agttagcaaa ggagaagaac ttttcactgg agttgtgcca attcttgttg aattagatgg
     1981 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg ctacatacgg
     2041 aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     2101 tgtcactact ctgacctatg gcgttcaatg cttttcccgt tatccggatc acatgaaacg
     2161 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
     2221 caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
     2281 taatcgtatc gagttaaagg gtattgattt taaagaagat ggaaacattc tcggacacaa
     2341 actagagtac aactataact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
     2401 aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggttccgttc aactagcaga
     2461 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     2521 cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct
     2581 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct acaaatgacc
     2641 aattattgaa ggccgctaac gcggcctttt tttgtttctg gtctcccaaa aatttatttg
     2701 cttattaatc atccggctcg tataatgtgt ggagggccca agttcactta aaaaggagaa
     2761 tagttatgaa atcgaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg
     2821 taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc gtccattccg
     2881 acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg caatttctat
     2941 gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt
     3001 cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc ctgtggattc
     3061 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     3121 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     3181 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctccc
     3241 tgcacgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     3301 tcctaatgca ggagtcgcat aagggagagc gccgtccgat gcccttgaga gccttcaacc
     3361 cagtcagctc cttccggtgg gcgcggggca tgaccattgt ggccgcactt atgactgtct
     3421 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     3481 accgctttcg ctggagcgcg acgatgagcg gcctgtcgct tgcggtattc ggaatcttgc
     3541 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     3601 ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc
     3661 gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg
     3721 cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat
     3781 cgctcgcggc tcttaccagc ctaacttcga tcactggacc gctgatcgtc acggcgattt
     3841 atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc
     3901 ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgagaca
     3961 attgtccaga taaaaaaaat ccttagcttt cgctaaggat gatttctaaa atttttctgt
     4021 catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt aaatcagact
     4081 gaagacttta tctctctgtc ataaaactgt catattcctt acatataact gtcacctgtt
     4141 tgtcctattt tgcttctcgt agccaacaaa caatgcttta tgaatcctcc caggagacat
     4201 tatgaaagtt atgcgtacca ccgtcgcaac tgttgtcgcc gcgaccttat cgatgagtgc
     4261 tttctctgtg tttgcagaag caagcctgac aggtgcaggt gcaaccttcc ctgcgccggt
     4321 gtatgccaaa tgggctgaca cttaccagaa agaaaccggt aataaagtta actaccaggg
     4381 tatcggttct tccggtggcg taaaacagat tatcgctaat accgttgatt ttggtgcctc
     4441 tgacgcgcc
//
