LOCUS       sequence-60470-e        3123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3123)
  AUTHORS   Reed B, Blazeck J, Alper H
  TITLE     Evolution of an alkane-inducible biosensor for increased 
            responsiveness to short-chain alkanes.
  JOURNAL   J Biotechnol. 2012 Apr 15;158(3):75-9. doi: 
            10.1016/j.jbiotec.2012.01.028. Epub 2012 Feb 2.
  PUBMED    22326628
REFERENCE   2  (bases 1 to 3123)
  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"; doi: 
            "10.1016/j.jbiotec.2012.01.028"; journalName: "J Biotechnol"; date: 
            "2012-04-15- 15"; volume: "158"; issue: "3"; pages: "75-9"
COMMENT     Sequence Label: pZE-PalkB-GFP
FEATURES             Location/Qualifiers
     source          1..3123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             306..317
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             326..1039
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             326..1039
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(356..384)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     979..999
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1142..1228
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1392..1980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1472..1491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2068..2162)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2193..2987)
                     /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(2304..2323)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2914..2933
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttac ccggaagccc gggttttgga ggtttccaat cgtgccttgg cgcagcgaca
       61 gccctcggtc ccccagatag ccattgatca ccgccagcat gcttgcggcc agctcgtgtt
      121 tttccagcag acgacggagc aaaaactacc cgtaggtgta gttggcgcaa gcgtccgatt
      181 agctcaggtt taagatgtcg agagtgagag tgggcggctt aactttctca gttaggcata
      241 aaattacgtc ttaaatctcg tagcgactaa tttaataaaa attggagaac accaaagaat
      301 tcattaaaga ggagaaaggt accgcatgcg taaaggagaa gaacttttca ctggagttgt
      361 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
      421 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
      481 actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc aatgctttgc
      541 gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
      601 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
      661 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
      721 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
      781 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga
      841 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
      901 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
      961 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     1021 catggatgaa ctatacaaaa ggcctgcagc aaacgacgaa aactacgctg cagcagttta
     1081 ataagcttga tatcgaattc ctgcagcccg ggggatccca tggtacgcgt gctagaggca
     1141 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1201 ggtgaacgct ctcctgagta ggacaaatcc gccgccctag acctaggcgt tcggctgcgg
     1261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1561 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     1621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1801 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     1861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2041 gggattttgg tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     2101 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     2161 tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     2221 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     2281 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     2341 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     2401 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     2461 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     2521 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     2581 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     2641 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     2701 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     2761 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     2821 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     2881 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     2941 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     3001 catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     3061 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     3121 tcc
//
