LOCUS       SEGA056                4448 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12892.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4448)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PT7-low-BCD-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4448
                     /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..1923
                     /codon_start=1
                     /label=BCD
                     /translation="MKAIFVLKHLNHAKEVF"
     misc_feature    1906..1914
                     /label=SD2
     misc_feature    1919..1921
                     /label=STOP BCD
     misc_feature    1921..1923
                     /label=start GOI
     CDS             1924..2634
                     /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    2638..2686
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2697..2725
                     /label=Promoter 3BCD2
     CDS             2765..3955
                     /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    3936..3985
                     /label=HR3
     terminator      3966..4006
                     /label=rrnC
     misc_feature    4007..4056
                     /label=homology
     CDS             4201..4448
                     /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 aggggggatg aaggctattt tcgtactgaa acatcttaat catgctaagg aggttttcta
     1921 atgagcaaag gagaagaact tttcactgga gttgtgccaa ttcttgttga attagatggt
     1981 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc tacatacgga
     2041 aaactcaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     2101 gtcactactc tgacctatgg cgttcaatgc ttttcccgtt atccggatca catgaaacgg
     2161 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
     2221 aaagatgacg ggaactacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
     2281 aatcgtatcg agttaaaggg tattgatttt aaagaagatg gaaacattct cggacacaaa
     2341 ctagagtaca actataactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
     2401 atcaaagcta acttcaaaat tcgccacaac attgaagatg gttccgttca actagcagac
     2461 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     2521 ctgtcgacac aatctgccct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     2581 cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta caaatgacca
     2641 attattgaag gccgctaacg cggccttttt ttgtttctgg tctcccaaaa atttatttgc
     2701 ttattaatca tccggctcgt ataatgtgtg gagggcccaa gttcacttaa aaaggagaat
     2761 agttatgaaa tcgaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt
     2821 aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga
     2881 cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg
     2941 cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc
     3001 gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc tgtggattct
     3061 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     3121 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     3181 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctccct
     3241 gcacgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     3301 cctaatgcag gagtcgcata agggagagcg ccgtccgatg cccttgagag ccttcaaccc
     3361 agtcagctcc ttccggtggg cgcggggcat gaccattgtg gccgcactta tgactgtctt
     3421 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     3481 ccgctttcgc tggagcgcga cgatgagcgg cctgtcgctt gcggtattcg gaatcttgca
     3541 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     3601 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     3661 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     3721 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     3781 gctcgcggct cttaccagcc taacttcgat cactggaccg ctgatcgtca cggcgattta
     3841 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     3901 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgagacaa
     3961 ttgtccagat aaaaaaaatc cttagctttc gctaaggatg atttctaaaa tttttctgtc
     4021 atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttattta aatcagactg
     4081 aagactttat ctctctgtca taaaactgtc atattcctta catataactg tcacctgttt
     4141 gtcctatttt gcttctcgta gccaacaaac aatgctttat gaatcctccc aggagacatt
     4201 atgaaagtta tgcgtaccac cgtcgcaact gttgtcgccg cgaccttatc gatgagtgct
     4261 ttctctgtgt ttgcagaagc aagcctgaca ggtgcaggtg caaccttccc tgcgccggtg
     4321 tatgccaaat gggctgacac ttaccagaaa gaaaccggta ataaagttaa ctaccagggt
     4381 atcggttctt ccggtggcgt aaaacagatt atcgctaata ccgttgattt tggtgcctct
     4441 gacgcgcc
//
