LOCUS       SEGA052                5246 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12888.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5246)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PRha-PelB-high-sfGFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..5246
                     /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..2721
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2722..3432
                     /codon_start=1
                     /label=sfGFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    3436..3484
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3495..3523
                     /label=Promoter 3BCD2
     CDS             3563..4753
                     /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    4734..4783
                     /label=HR3
     terminator      4764..4804
                     /label=rrnC
     misc_feature    4805..4854
                     /label=homology
     CDS             4999..5246
                     /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 gaggagaaat taattatgaa atatctgctg ccgaccgctg ctgctggtct gctgctcctc
     2701 gctgcccagc cggcgatggc ccgtaaaggc gaagagctgt tcactggtgt cgtccctatt
     2761 ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa
     2821 ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg
     2881 gtaccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat
     2941 ccggaccaca tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag
     3001 gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt
     3061 gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaagacggc
     3121 aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc
     3181 gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc
     3241 agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg
     3301 ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa
     3361 cgcgatcaca tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat
     3421 gaactgtaca aatgaccaat tattgaaggc cgctaacgcg gccttttttt gtttctggtc
     3481 tcccaaaaat ttatttgctt attaatcatc cggctcgtat aatgtgtgga gggcccaagt
     3541 tcacttaaaa aggagaatag ttatgaaatc gaacaatgcg ctcatcgtca tcctcggcac
     3601 cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg
     3661 ggatatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc
     3721 gttgatgcaa tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg
     3781 cccagtcctg ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac
     3841 acccgtcctg tggattctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     3901 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     3961 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     4021 gttgggcgcc atctccctgc acgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     4081 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgcc gtccgatgcc
     4141 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ccattgtggc
     4201 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg
     4261 ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgagcggcc tgtcgcttgc
     4321 ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg
     4381 tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt
     4441 gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg
     4501 cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca
     4561 gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct
     4621 gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt
     4681 aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc
     4741 cacctcgacc tgagacaatt gtccagataa aaaaaatcct tagctttcgc taaggatgat
     4801 ttctaaaatt tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg
     4861 cttatttaaa tcagactgaa gactttatct ctctgtcata aaactgtcat attccttaca
     4921 tataactgtc acctgtttgt cctattttgc ttctcgtagc caacaaacaa tgctttatga
     4981 atcctcccag gagacattat gaaagttatg cgtaccaccg tcgcaactgt tgtcgccgcg
     5041 accttatcga tgagtgcttt ctctgtgttt gcagaagcaa gcctgacagg tgcaggtgca
     5101 accttccctg cgccggtgta tgccaaatgg gctgacactt accagaaaga aaccggtaat
     5161 aaagttaact accagggtat cggttcttcc ggtggcgtaa aacagattat cgctaatacc
     5221 gttgattttg gtgcctctga cgcgcc
//
