LOCUS       SEGA075                4423 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12911.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4423)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PSal-high-BCD-GFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..4423
                     /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(611..1513)
                     /label=NahR-AM
     misc_feature    770..772
                     /label=R425C
                     /note="Geneious type: mutation"
     misc_feature    1160..1162
                     /label=V118A
                     /note="Geneious type: mutation"
     misc_feature    1298..1300
                     /label=P72H
                     /note="Geneious type: mutation"
     misc_feature    1439..1441
                     /label=I25V
                     /note="Geneious type: mutation"
     RBS             1514..1542
                     /label=nah2
     misc_feature    1543..1596
                     /label=pLacIQ
                     /note="Geneious type: promoter"
     promoter        1550..1555
                     /label=-10
     promoter        1573..1578
                     /label=-35
     promoter        1623..1738
                     /label=pSalTTC
     misc_feature    1660..1690
                     /label=NahO
                     /note="Geneious type: operator"
     promoter        1704..1709
                     /label=-35
     promoter        1726..1731
                     /label=-10
     misc_feature    1739..1813
                     /label=RiboJ
                     /note="Geneious type: insulator"
     RBS             1825..1836
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    1837..1851
                     /label=TIR
     CDS             1843..1898
                     /codon_start=1
                     /label=BCD
                     /translation="MKAIFVLKHLNHAKEVF"
     misc_feature    1881..1889
                     /label=SD2
     misc_feature    1894..1896
                     /label=STOP BCD
     misc_feature    1896..1898
                     /label=start GOI
     CDS             1899..2609
                     /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    2613..2661
                     /label=L3S3P22 Terminator (synthetic)
     promoter        2672..2700
                     /label=Promoter 3BCD2
     CDS             2740..3930
                     /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    3911..3960
                     /label=HR3
     terminator      3941..3981
                     /label=rrnC
     misc_feature    3982..4031
                     /label=homology
     CDS             4176..4423
                     /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 tcaatccgta aacaggtcaa acatcagttg ccgcaaccaa atattggcta
      661 ggtccttgtg gtacttcgca tgccagaaca tgttgatggc tatttcaggc aagacgactg
      721 ggtgcggcaa ggcgcttagg ccgaagggct ctacgcagca gtcggctaaa catatcggca
      781 cagtggcgag cagatcggtg cgctggagga tgtggccaac ggcggcgaag tgcggcactt
      841 ccagacggat gtcgcgccgg atgccgaccc gtgtcatgta cgtgtccacc tcgccgtggc
      901 cggtgccagc ggcgatgaca cgcacgtggc cgtaggaaca gaagcgctcc agagtcaggg
      961 gttcgcgggt gactggatgg tccttgcgac ataggcacac gtagtgattc tggagcagcc
     1021 ggcgctgaaa gaagccagtt tgcagattgg gaagcaggcc cacggccaag tccacggttc
     1081 cgttctgcaa ggcctgcatc aggctcatcg aactgtcgcg caccgtactg atcacgcaat
     1141 tgggggcctg gtgagccagc gcatccatca gccgcggcat gaagtagatc tcgccaatgt
     1201 cggtcatggc cagggtgaag gtacgctcgc tggtcagcgg atcgaagctt tcatggtgct
     1261 gtagggcgtt gcgcagtgcg tgcatggccg aagtgacgtg ctcggccaga tgcgcggcat
     1321 agggtgtggg ttccattccc tgatgtgtgc gcacgaagag tgggtcctgt agcgaggtgc
     1381 gcaggcgttt cagcgcattg ctcacggcag gctgggtcag gcccaggttc tccgcagtga
     1441 cagagacgcg tctgtcgacc agcaactggt tgaacaccac cagcaggttt aaatccaggt
     1501 cacgcagttc catattcaac ccccatgaat tgactctctt ccgggcgcta tcatgccata
     1561 ccgcgaaagg ttttgcacca ttcgatggcg cgccgccatt ggagcttaac gatcgttggc
     1621 tgggggcctc gcttgggtta ttgctggtgc ccggccgggc gcaatattca tgttgatgat
     1681 ttattatata tcgagtggtg tatttattta tattgtttgc tccgttaccg ttattaacag
     1741 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
     1801 taattttgtt taatactaga gattaaagag gagaaactat atatgaaagc aattttcgta
     1861 ctgaaacatc ttaatcatgc taaggaggtt ttctaatgag caaaggagaa gaacttttca
     1921 ctggagttgt gccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     1981 tcagtggaga gggtgaaggt gatgctacat acggaaaact cacccttaaa tttatttgca
     2041 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactctgacc tatggcgttc
     2101 aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc
     2161 ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggaac tacaagacgc
     2221 gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg
     2281 attttaaaga agatggaaac attctcggac acaaactaga gtacaactat aactcacaca
     2341 atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc
     2401 acaacattga agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg
     2461 gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaatct gccctttcga
     2521 aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga
     2581 ttacacatgg catggatgag ctctacaaat gaccaattat tgaaggccgc taacgcggcc
     2641 tttttttgtt tctggtctcc caaaaattta tttgcttatt aatcatccgg ctcgtataat
     2701 gtgtggaggg cccaagttca cttaaaaagg agaatagtta tgaaatcgaa caatgcgctc
     2761 atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta
     2821 ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg
     2881 ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc
     2941 gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac
     3001 gcgatcatgg cgaccacacc cgtcctgtgg attctctacg ccggacgcat cgtggccggc
     3061 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     3121 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     3181 cccgtggccg ggggactgtt gggcgccatc tccctgcacg caccattcct tgcggcggcg
     3241 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     3301 gagcgccgtc cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3361 ggcatgacca ttgtggccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3421 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     3481 agcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     3541 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
     3601 gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
     3661 attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
     3721 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact
     3781 tcgatcactg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
     3841 gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
     3901 ggtgcatgga gccgggccac ctcgacctga gacaattgtc cagataaaaa aaatccttag
     3961 ctttcgctaa ggatgatttc taaaattttt ctgtcatctc ttcgttatta atgtttgtaa
     4021 ttgactgaat atcaacgctt atttaaatca gactgaagac tttatctctc tgtcataaaa
     4081 ctgtcatatt ccttacatat aactgtcacc tgtttgtcct attttgcttc tcgtagccaa
     4141 caaacaatgc tttatgaatc ctcccaggag acattatgaa agttatgcgt accaccgtcg
     4201 caactgttgt cgccgcgacc ttatcgatga gtgctttctc tgtgtttgca gaagcaagcc
     4261 tgacaggtgc aggtgcaacc ttccctgcgc cggtgtatgc caaatgggct gacacttacc
     4321 agaaagaaac cggtaataaa gttaactacc agggtatcgg ttcttccggt ggcgtaaaac
     4381 agattatcgc taataccgtt gattttggtg cctctgacgc gcc
//
