LOCUS       SEGA077                4103 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12913.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4103)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PTet-high-BCD-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4103
                     /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
     misc_feature    complement(611..1234)
                     /label=TetR
                     /note="Geneious type: CDS"
     RBS             1235..1262
                     /label=tet1
     misc_feature    1263..1316
                     /label=pLacI
                     /note="Geneious type: promoter"
     promoter        1270..1275
                     /label=-10
     promoter        1293..1298
                     /label=-35
     promoter        1343..1418
                     /label=pTet*
     misc_feature    1365..1383
                     /label=TetO2
                     /note="Geneious type: operator"
     promoter        1384..1389
                     /label=-35
     misc_feature    1390..1408
                     /label=TetO2
                     /note="Geneious type: operator"
     promoter        1407..1412
                     /label=-10
     misc_feature    1419..1493
                     /label=RiboJ
                     /note="Geneious type: insulator"
     RBS             1505..1516
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    1517..1531
                     /label=TIR
     CDS             1523..1578
                     /codon_start=1
                     /label=BCD
                     /translation="MKAIFVLKHLNHAKEVF"
     misc_feature    1561..1569
                     /label=SD2
     misc_feature    1574..1576
                     /label=STOP BCD
     misc_feature    1576..1578
                     /label=start GOI
     CDS             1579..2289
                     /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    2293..2341
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2352..2380
                     /label=Promoter 3BCD2
     CDS             2420..3610
                     /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    3591..3640
                     /label=HR3
     terminator      3621..3661
                     /label=rrnC
     misc_feature    3662..3711
                     /label=homology
     CDS             3856..4103
                     /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 gtccccaatt
      601 aggatcctta tcaggaccca ctttcacatt taagttgttt ttctaatccg catatgatca
      661 attcaaggcc gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt
      721 gtcgtaataa tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt
      781 gatgctcttg atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat
      841 ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag
      901 tttcatactg tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac
      961 ttagtaaagc acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc
     1021 cttctaaagg gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga
     1081 gcaaagcccg cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct
     1141 gggcgagttt acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc
     1201 tgttaatcac tttactttta tctaatctgg acatattcac caccctgaat tgactctctt
     1261 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggcg cgccgccatt
     1321 ggagcttaac gatcgttggc tgttttcagc aggacgcact gacctcccta tcagtgatag
     1381 agattgacat ccctatcagt gatagagata ctgagcacag ctgtcaccgg atgtgctttc
     1441 cggtctgatg agtccgtgag gacgaaacag cctctacaaa taattttgtt taatactaga
     1501 gattaaagag gagaaactat atatgaaagc aattttcgta ctgaaacatc ttaatcatgc
     1561 taaggaggtt ttctaatgag caaaggagaa gaacttttca ctggagttgt gccaattctt
     1621 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
     1681 gatgctacat acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
     1741 ccatggccaa cacttgtcac tactctgacc tatggcgttc aatgcttttc ccgttatccg
     1801 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     1861 cgcactatat ctttcaaaga tgacgggaac tacaagacgc gtgctgaagt caagtttgaa
     1921 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
     1981 attctcggac acaaactaga gtacaactat aactcacaca atgtatacat cacggcagac
     2041 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacattga agatggttcc
     2101 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     2161 ccagacaacc attacctgtc gacacaatct gccctttcga aagatcccaa cgaaaagcgt
     2221 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     2281 ctctacaaat gaccaattat tgaaggccgc taacgcggcc tttttttgtt tctggtctcc
     2341 caaaaattta tttgcttatt aatcatccgg ctcgtataat gtgtggaggg cccaagttca
     2401 cttaaaaagg agaatagtta tgaaatcgaa caatgcgctc atcgtcatcc tcggcaccgt
     2461 caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga
     2521 tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt
     2581 gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc
     2641 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
     2701 cgtcctgtgg attctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     2761 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     2821 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     2881 gggcgccatc tccctgcacg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     2941 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgccgtc cgatgccctt
     3001 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacca ttgtggccgc
     3061 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     3121 cattttcggc gaggaccgct ttcgctggag cgcgacgatg agcggcctgt cgcttgcggt
     3181 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     3241 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct
     3301 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg
     3361 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg
     3421 acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg gaccgctgat
     3481 cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg
     3541 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     3601 ctcgacctga gacaattgtc cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     3661 taaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat atcaacgctt
     3721 atttaaatca gactgaagac tttatctctc tgtcataaaa ctgtcatatt ccttacatat
     3781 aactgtcacc tgtttgtcct attttgcttc tcgtagccaa caaacaatgc tttatgaatc
     3841 ctcccaggag acattatgaa agttatgcgt accaccgtcg caactgttgt cgccgcgacc
     3901 ttatcgatga gtgctttctc tgtgtttgca gaagcaagcc tgacaggtgc aggtgcaacc
     3961 ttccctgcgc cggtgtatgc caaatgggct gacacttacc agaaagaaac cggtaataaa
     4021 gttaactacc agggtatcgg ttcttccggt ggcgtaaaac agattatcgc taataccgtt
     4081 gattttggtg cctctgacgc gcc
//
