LOCUS       SEGA039                5236 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12875.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5236)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PRha-medium-BCD-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..5236
                     /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..2711
                     /codon_start=1
                     /label=BCD
                     /translation="MKAIFVLKHLNHAKEVF"
     misc_feature    2694..2702
                     /label=SD2
     misc_feature    2707..2709
                     /label=STOP BCD
     misc_feature    2709..2711
                     /label=start GOI
     CDS             2712..3422
                     /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    3426..3474
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3485..3513
                     /label=Promoter 3BCD2
     CDS             3553..4743
                     /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    4724..4773
                     /label=HR3
     terminator      4754..4794
                     /label=rrnC
     misc_feature    4795..4844
                     /label=homology
     CDS             4989..5236
                     /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 gaggagaaag agctaatgaa ggctattttc gtactgaaac atcttaatca tgctaaggag
     2701 gttttctaat gagcaaagga gaagaacttt tcactggagt tgtgccaatt cttgttgaat
     2761 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcta
     2821 catacggaaa actcaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     2881 caacacttgt cactactctg acctatggcg ttcaatgctt ttcccgttat ccggatcaca
     2941 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta
     3001 tatctttcaa agatgacggg aactacaaga cgcgtgctga agtcaagttt gaaggtgata
     3061 cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattctcg
     3121 gacacaaact agagtacaac tataactcac acaatgtata catcacggca gacaaacaaa
     3181 agaatggaat caaagctaac ttcaaaattc gccacaacat tgaagatggt tccgttcaac
     3241 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     3301 accattacct gtcgacacaa tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca
     3361 tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca
     3421 aatgaccaat tattgaaggc cgctaacgcg gccttttttt gtttctggtc tcccaaaaat
     3481 ttatttgctt attaatcatc cggctcgtat aatgtgtgga gggcccaagt tcacttaaaa
     3541 aggagaatag ttatgaaatc gaacaatgcg ctcatcgtca tcctcggcac cgtcaccctg
     3601 gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc
     3661 cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa
     3721 tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg
     3781 ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg
     3841 tggattctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     3901 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     3961 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     4021 atctccctgc acgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     4081 ggctgcttcc taatgcagga gtcgcataag ggagagcgcc gtccgatgcc cttgagagcc
     4141 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ccattgtggc cgcacttatg
     4201 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     4261 ggcgaggacc gctttcgctg gagcgcgacg atgagcggcc tgtcgcttgc ggtattcgga
     4321 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     4381 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     4441 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     4501 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     4561 caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg
     4621 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     4681 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     4741 tgagacaatt gtccagataa aaaaaatcct tagctttcgc taaggatgat ttctaaaatt
     4801 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttaaa
     4861 tcagactgaa gactttatct ctctgtcata aaactgtcat attccttaca tataactgtc
     4921 acctgtttgt cctattttgc ttctcgtagc caacaaacaa tgctttatga atcctcccag
     4981 gagacattat gaaagttatg cgtaccaccg tcgcaactgt tgtcgccgcg accttatcga
     5041 tgagtgcttt ctctgtgttt gcagaagcaa gcctgacagg tgcaggtgca accttccctg
     5101 cgccggtgta tgccaaatgg gctgacactt accagaaaga aaccggtaat aaagttaact
     5161 accagggtat cggttcttcc ggtggcgtaa aacagattat cgctaatacc gttgattttg
     5221 gtgcctctga cgcgcc
//
