LOCUS       SEGA043                5489 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12879.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5489)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PRha-high-Ubiquitin-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..5489
                     /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..2955
                     /codon_start=1
                     /label=Ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPGSAHHHHHHHHHHAGSSDTIENVKAK
                     IQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQEKSTLELVLELQNETQSATSDA"
     CDS             2722..2751
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2965..3675
                     /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    3679..3727
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3738..3766
                     /label=Promoter 3BCD2
     CDS             3806..4996
                     /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    4977..5026
                     /label=HR3
     terminator      5007..5047
                     /label=rrnC
     misc_feature    5048..5097
                     /label=homology
     CDS             5242..5489
                     /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 attgtatgca gatattcgtg aaaacactga ccggcaaaac cattaccctg
     2701 gaggtggagc ctggcagcgc ccatcatcat caccatcacc atcaccatca tgctggtagc
     2761 agcgatacga tcgaaaacgt caaagccaaa attcaggaca aagagggcat tccgcctgat
     2821 caacaacgtc tgatcttcgc cggaaaacag ctggaggatg gccgtacact gagtgactat
     2881 aacattcaag agaaatcgac actggaactg gttctggagc tgcagaatga aacccagtca
     2941 gcaacttctg acgcttctgg cggtagcaaa ggagaagaac ttttcactgg agttgtgcca
     3001 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     3061 gaaggtgatg ctacatacgg aaaactcacc cttaaattta tttgcactac tggaaaacta
     3121 cctgttccat ggccaacact tgtcactact ctgacctatg gcgttcaatg cttttcccgt
     3181 tatccggatc acatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     3241 caggaacgca ctatatcttt caaagatgac gggaactaca agacgcgtgc tgaagtcaag
     3301 tttgaaggtg atacccttgt taatcgtatc gagttaaagg gtattgattt taaagaagat
     3361 ggaaacattc tcggacacaa actagagtac aactataact cacacaatgt atacatcacg
     3421 gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cattgaagat
     3481 ggttccgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     3541 cttttaccag acaaccatta cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa
     3601 aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg
     3661 gatgagctct acaaatgacc aattattgaa ggccgctaac gcggcctttt tttgtttctg
     3721 gtctcccaaa aatttatttg cttattaatc atccggctcg tataatgtgt ggagggccca
     3781 agttcactta aaaaggagaa tagttatgaa atcgaacaat gcgctcatcg tcatcctcgg
     3841 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     3901 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
     3961 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
     4021 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
     4081 cacacccgtc ctgtggattc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     4141 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     4201 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     4261 actgttgggc gccatctccc tgcacgcacc attccttgcg gcggcggtgc tcaacggcct
     4321 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gccgtccgat
     4381 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgaccattgt
     4441 ggccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     4501 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgagcg gcctgtcgct
     4561 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     4621 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     4681 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     4741 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     4801 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc
     4861 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     4921 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     4981 ggccacctcg acctgagaca attgtccaga taaaaaaaat ccttagcttt cgctaaggat
     5041 gatttctaaa atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca
     5101 acgcttattt aaatcagact gaagacttta tctctctgtc ataaaactgt catattcctt
     5161 acatataact gtcacctgtt tgtcctattt tgcttctcgt agccaacaaa caatgcttta
     5221 tgaatcctcc caggagacat tatgaaagtt atgcgtacca ccgtcgcaac tgttgtcgcc
     5281 gcgaccttat cgatgagtgc tttctctgtg tttgcagaag caagcctgac aggtgcaggt
     5341 gcaaccttcc ctgcgccggt gtatgccaaa tgggctgaca cttaccagaa agaaaccggt
     5401 aataaagtta actaccaggg tatcggttct tccggtggcg taaaacagat tatcgctaat
     5461 accgttgatt ttggtgcctc tgacgcgcc
//
