LOCUS       sequence-197084-        5192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The pAK plasmid expresses GFP driven by a T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  AUTHORS   Kushwaha M, Salis HM
  TITLE     A portable expression resource for engineering cross-species genetic
            circuits and pathways.
  JOURNAL   Nat Commun. 2015 Jul 17;6:7832. doi: 10.1038/ncomms8832.
  PUBMED    26184393
REFERENCE   2  (bases 1 to 5192)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2015-07-17"; volume: "6:7832. doi"; pages: " 
            10.1038/ncomms8832"
COMMENT     Sequence Label: MK969
FEATURES             Location/Qualifiers
     source          1..5192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(617..1411)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(728..747)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1338..1357
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        1612..1630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1652..1660
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             join(1678..1680,1681..1683,1684..2394)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      2410..2457
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3051..3713)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLSGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3714..4485
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca
       61 ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc
      121 ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg
      181 cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga
      241 gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca
      301 agagacggcc cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt
      361 gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct
      421 gacggaacag cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg
      481 cgcacatttc cccgaaaagt gccacctgga ttttggtcat gagattatca aaaaggatct
      541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      601 aaacttggtc tgacagtcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
      661 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
      721 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
      781 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
      841 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
      901 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
      961 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     1021 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     1081 cggcaggagc aaggtgggat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     1141 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     1201 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     1261 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     1321 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     1381 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     1441 cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     1501 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     1561 tccataaaac cgcccagtct agctatcgcc atgacctagt ggatccgaaa ttaatacgac
     1621 tcactatagg aggattatat aaacagcgat ttaaggaggt aaaaaatggg cgacgtcatg
     1681 gtttcaaagg gtgaagaatt atttacaggt gttgttccta ttttagttga attagatggt
     1741 gatgttaatg gtcataagtt ctcagtttca ggtgaaggtg aaggtgatgc aacatatggt
     1801 aagttaacat taaagttcat ttgtacaaca ggtaaattac ctgttccttg gcctacatta
     1861 gttacaacat taacatatgg tgttcaatgt ttttcacgtt atcctgatca tatgaaacaa
     1921 catgacttct ttaaatcagc aatgccagaa ggttacgttc aagaacgtac catctttttt
     1981 aaagatgatg gtaactataa aacacgtgca gaagttaagt tcgaaggtga tacattagtt
     2041 aatcgtattg aattaaaagg tattgacttc aaagaagatg gtaatatttt aggtcataaa
     2101 ttagaatata actataattc acataatgtt tatattatgg cagataaaca gaaaaatggt
     2161 attaaagtta acttcaagat acgtcataat attgaagatg gttcagttca gttagcagat
     2221 cattatcaac agaatactcc tattggtgat ggtcctgttt tattacctga taatcattac
     2281 ttatcaaccc aatcagcatt atcaaaagat cctaatgaaa aacgtgatca tatggtttta
     2341 ttagagttcg ttacagcagc aggtattaca ttaggtatgg atgaattata taaatgataa
     2401 tctagataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2461 cctagcgatg aatggcgaat ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     2521 aatttttgtt aaatcagctc attttttaac caataggccg tactgcgatg agtggcaggg
     2581 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     2641 aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc
     2701 ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg
     2761 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     2821 ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca gagcctgata
     2881 aaaacggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc
     2941 tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg aggcggctac
     3001 agccgatagt ctggaacagc gcacttacgg gttgctgcgc aacccaagtg ctaccggcgc
     3061 ggcagcgtga cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa acacggctca
     3121 tcgggcatcg gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct
     3181 ggcaggtctg gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg
     3241 gtcggtagtt gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc
     3301 aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg
     3361 cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg
     3421 gtgccggggc cgctgagctt cacgacggag atccagcgct cggccaccaa gtccttgact
     3481 gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagtgtctt ctggctgacc
     3541 accacggcgt tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg
     3601 ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag
     3661 tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc
     3721 tccatgcggt aggggtgccg cacggttgcg gcaccatgcg caatcagctg caacttttcg
     3781 gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt caattacaga ttttctttaa
     3841 cctacgcaat gagctattgc ggggggtgcc gcaatgagct gttgcgtacc cccctttttt
     3901 aagttgttga tttttaagtc tttcgcattt cgccctatat ctagttcttt ggtgcccaaa
     3961 gaagggcacc cctgcggggt tcccccacgc cttcggcgcg gctccccctc cggcaaaaag
     4021 tggcccctcc ggggcttgtt gatcgactgc gcggccttcg gccttgccca aggtggcgct
     4081 gcccccttgg aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc
     4141 gcccttcgac tgcccccact cgcataggct tgggtcgttc caggcgcgtc aaggccaagc
     4201 cgctgcgcgg tcgctgcgcg agccttgacc cgccttccac ttggtgtcca accggcaagc
     4261 gaagcgcgca ggccgcaggc cggaggcttt tccccagaga aaattaaaaa aattgatggg
     4321 gcaaggccgc aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg
     4381 gtgggcaatc cctgtggtca agctcgtggg caggcgcagc ctgtccatca gcttgtccag
     4441 cagggttgtc cacgggccga gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc
     4501 acatatccac gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa
     4561 gcccgtaggg cgccgcagcc gccgtaggcg gtcacgactt tgcgaagcaa agtctagtga
     4621 gtatactcaa gcgttgagtg gcccgccgga ggcaccgcct tgcgctgccc ccgtcgagcc
     4681 ggttggacac caaaagggag gggcaggcat ggcggcatac gcgatcatgc gatgcaagaa
     4741 gctggcgaaa atgggcaacg tggcggccag tctcaagcac gcctaccgcg agcgcgagac
     4801 gccaacgctg acgccagcag gacgccagag aacgagcact gggcggccag cagcaccgat
     4861 gaagcgatgg gccgactgcg cgagttgctg ccagagaagc ggcgcaagga cgctgtgttg
     4921 gcggtcgagt acgtcatgac ggccagcccg gaatggtgga agtcggccag ccaagaacag
     4981 caggcggcgt tcttcgagaa ggcgcacaag tggctggcgg acaagtacgg ggcggatcgc
     5041 atcgtgacgg ccagcatcca ccgtgacgaa accagcccgc acatgaccgc gttcgtggtg
     5101 ccgctgacgc aggacggcag gctgtcggcc aaggagttca tcggcaacaa agcgcagatg
     5161 acccgcgacc agaccacgtt tgcggccgct gt
//
