LOCUS       SEGA049                5237 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12885.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5237)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PRha-high-PROTi-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..5237
                     /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(607..1452)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(1526..2362)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter"
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        2508..2626
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     RBS             2638..2649
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    2650..2664
                     /label=TIR
     CDS             2656..2676
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="MENLYFQ"
     misc_feature    2677..2712
                     /label=PROTi-Linker
     CDS             2713..3423
                     /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    3427..3475
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3486..3514
                     /label=Promoter 3BCD2
     CDS             3554..4744
                     /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    4725..4774
                     /label=HR3
     terminator      4755..4795
                     /label=rrnC
     misc_feature    4796..4845
                     /label=homology
     CDS             4990..5237
                     /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 cgcttaatct ttctgcgaat tgagatgacg ccactggctg ggcgtcatcc cggtttcccg
      661 ggtaaacacc accgaaaaat agttactatc ttcaaagcca cattcggtcg aaatatcact
      721 gattaacagg cggctatgct ggagaagata ttgcgcatga cacactctga cctgtcgcag
      781 atattgattg atggtcattc cagtctgctg gcgaaattgc tgacgcaaaa cgcgctcact
      841 gcacgatgcc tcatcacaaa atttatccag cgcaaaggga cttttcaggc tagccgccag
      901 ccgggtaatc agcttatcca gcaacgtttc gctggatgtt ggcggcaacg aatcactggt
      961 gtaacgatgg cgattcagca acatcaccaa ctgcccgaac agcaactcag ccatttcgtt
     1021 agcaaacggc acatgctgac tactttcatg ctcaagctga ccaataacct gccgcgcctg
     1081 cgccatcccc atgctaccta agcgccagtg tggttgccct gcgctggcgt taaatcccgg
     1141 aatcgccccc tgccagtcaa gattcagctt cagacgctcc gggcaataaa taatattctg
     1201 caaaaccaga tcgttaacgg aagcgtagga gtgtttatca tcagcatgaa tgtaaaagag
     1261 atcgccacgg gtaatgcgat aagggcgatc gttgagtaca tgcaggccat taccgcgcca
     1321 gacaatcacc agctcacaaa aatcatgtgt atgttcagca aagacatctt gcggataacg
     1381 gtcagccaca gcgactgcct gctggtcgct ggcaaaaaaa tcatctttga gaagttttaa
     1441 ctgatgcgcc accgtggcta cctcggccag agaacgaagt tgattattcg caatatggcg
     1501 tacaaatacg ttgagaagat tcgcgttatt gcagaaagcc atcccgtccc tggcgaatat
     1561 cacgcggtga ccagttaaac tctcggcgaa aaagcgtcga aaagtggtta ctgtcgctga
     1621 atccacagcg ataggcgatg tcagtaacgc tggcctcgct gtggcgtagc agatgtcggg
     1681 ctttcatcag tcgcaggcgg ttcaggtatc gctgaggcgt cagtcccgtt tgctgcttaa
     1741 gctgccgatg tagcgtacgc agtgaaagag aaaattgatc cgccacggca tcccaattca
     1801 cctcatcggc aaaatggtcc tccagccagg ccagaagcaa gttgagacgt gatgcgctgt
     1861 tttccaggtt ctcctgcaaa ctgcttttac gcagcaagag cagtaattgc ataaacaaga
     1921 tctcgcgact ggcggtcgag ggtaaatcat tttccccttc ctgctgttcc atctgtgcaa
     1981 ccagctgtcg cacctgctgc aatacgctgt ggttaacgcg ccagtgagac ggatactgcc
     2041 catccagctc ttgtggcagc aactgattca gcccggcgag aaactgaaat cgatccggcg
     2101 agcgatacag cacattggtc agacacagat tatcggtatg ttcatacaga tgccgatcat
     2161 gatcgcgtac gaaacagacc gtgccaccgg tgatggtata gggctgccca ttaaacacat
     2221 gaatacccgt gccatgttcg acaatcacaa tttcatgaaa atcatgatga tgttcaggaa
     2281 aatccgcctg cgggagccgg ggttctatcg ccacggacgc gttaccagac ggaaaaaaat
     2341 ccacactatg taatacggtc atactggcct cctgatgtcg tcaacacggc gaaatagtaa
     2401 tcacgaggtc aggttcttac cttaaatttt cgacggaaaa ccacgtaaaa aacgtcgatt
     2461 tttcaagata cagcgtgaat tttcaggaaa tgcggtgagc atcacatcac cacaattcag
     2521 caaattgtga acatcatcac gttcatcttt ccctggttgc caatggccca ttttcttgtc
     2581 agtaacgaga aggtcgcgaa tccaggcgct ttttagactg gtcgtatact agagattaaa
     2641 gaggagaaaa tatcgatgga gaacctgtac ttccagtttc gtagcaaagg agaagaactt
     2701 gtcactggag ttagcaaagg agaagaactt ttcactggag ttgtgccaat tcttgttgaa
     2761 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgct
     2821 acatacggaa aactcaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     2881 ccaacacttg tcactactct gacctatggc gttcaatgct tttcccgtta tccggatcac
     2941 atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact
     3001 atatctttca aagatgacgg gaactacaag acgcgtgctg aagtcaagtt tgaaggtgat
     3061 acccttgtta atcgtatcga gttaaagggt attgatttta aagaagatgg aaacattctc
     3121 ggacacaaac tagagtacaa ctataactca cacaatgtat acatcacggc agacaaacaa
     3181 aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg ttccgttcaa
     3241 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     3301 aaccattacc tgtcgacaca atctgccctt tcgaaagatc ccaacgaaaa gcgtgaccac
     3361 atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac
     3421 aaatgaccaa ttattgaagg ccgctaacgc ggcctttttt tgtttctggt ctcccaaaaa
     3481 tttatttgct tattaatcat ccggctcgta taatgtgtgg agggcccaag ttcacttaaa
     3541 aaggagaata gttatgaaat cgaacaatgc gctcatcgtc atcctcggca ccgtcaccct
     3601 ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt
     3661 ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca
     3721 atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct
     3781 gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct
     3841 gtggattctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     3901 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     3961 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
     4021 catctccctg cacgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
     4081 gggctgcttc ctaatgcagg agtcgcataa gggagagcgc cgtccgatgc ccttgagagc
     4141 cttcaaccca gtcagctcct tccggtgggc gcggggcatg accattgtgg ccgcacttat
     4201 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     4261 cggcgaggac cgctttcgct ggagcgcgac gatgagcggc ctgtcgcttg cggtattcgg
     4321 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     4381 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt
     4441 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     4501 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     4561 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc tgatcgtcac
     4621 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     4681 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     4741 ctgagacaat tgtccagata aaaaaaatcc ttagctttcg ctaaggatga tttctaaaat
     4801 ttttctgtca tctcttcgtt attaatgttt gtaattgact gaatatcaac gcttatttaa
     4861 atcagactga agactttatc tctctgtcat aaaactgtca tattccttac atataactgt
     4921 cacctgtttg tcctattttg cttctcgtag ccaacaaaca atgctttatg aatcctccca
     4981 ggagacatta tgaaagttat gcgtaccacc gtcgcaactg ttgtcgccgc gaccttatcg
     5041 atgagtgctt tctctgtgtt tgcagaagca agcctgacag gtgcaggtgc aaccttccct
     5101 gcgccggtgt atgccaaatg ggctgacact taccagaaag aaaccggtaa taaagttaac
     5161 taccagggta tcggttcttc cggtggcgta aaacagatta tcgctaatac cgttgatttt
     5221 ggtgcctctg acgcgcc
//
