LOCUS       SEGA044                5201 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12880.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5201)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PRha-low-TEV-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..5201
                     /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"
     CDS             2677..3387
                     /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    3391..3439
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3450..3478
                     /label=Promoter 3BCD2
     CDS             3518..4708
                     /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    4689..4738
                     /label=HR3
     terminator      4719..4759
                     /label=rrnC
     misc_feature    4760..4809
                     /label=homology
     CDS             4954..5201
                     /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 gaggagaaac gggggatgga gaacctgtac ttccagagca aaggagaaga acttttcact
     2701 ggagttgtgc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     2761 agtggagagg gtgaaggtga tgctacatac ggaaaactca cccttaaatt tatttgcact
     2821 actggaaaac tacctgttcc atggccaaca cttgtcacta ctctgaccta tggcgttcaa
     2881 tgcttttccc gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc
     2941 gaaggttatg tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt
     3001 gctgaagtca agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat
     3061 tttaaagaag atggaaacat tctcggacac aaactagagt acaactataa ctcacacaat
     3121 gtatacatca cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac
     3181 aacattgaag atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc
     3241 gatggccctg tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa
     3301 gatcccaacg aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt
     3361 acacatggca tggatgagct ctacaaatga ccaattattg aaggccgcta acgcggcctt
     3421 tttttgtttc tggtctccca aaaatttatt tgcttattaa tcatccggct cgtataatgt
     3481 gtggagggcc caagttcact taaaaaggag aatagttatg aaatcgaaca atgcgctcat
     3541 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
     3601 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct
     3661 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga
     3721 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
     3781 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat
     3841 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     3901 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     3961 cgtggccggg ggactgttgg gcgccatctc cctgcacgca ccattccttg cggcggcggt
     4021 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     4081 gcgccgtccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     4141 catgaccatt gtggccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     4201 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgag
     4261 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     4321 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     4381 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     4441 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     4501 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     4561 gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     4621 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     4681 tgcatggagc cgggccacct cgacctgaga caattgtcca gataaaaaaa atccttagct
     4741 ttcgctaagg atgatttcta aaatttttct gtcatctctt cgttattaat gtttgtaatt
     4801 gactgaatat caacgcttat ttaaatcaga ctgaagactt tatctctctg tcataaaact
     4861 gtcatattcc ttacatataa ctgtcacctg tttgtcctat tttgcttctc gtagccaaca
     4921 aacaatgctt tatgaatcct cccaggagac attatgaaag ttatgcgtac caccgtcgca
     4981 actgttgtcg ccgcgacctt atcgatgagt gctttctctg tgtttgcaga agcaagcctg
     5041 acaggtgcag gtgcaacctt ccctgcgccg gtgtatgcca aatgggctga cacttaccag
     5101 aaagaaaccg gtaataaagt taactaccag ggtatcggtt cttccggtgg cgtaaaacag
     5161 attatcgcta ataccgttga ttttggtgcc tctgacgcgc c
//
