LOCUS       sequence-60471-e        6646 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 6646)
  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 6646)
  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: pACYC-PLlambda-alkS(WT)
FEATURES             Location/Qualifiers
     source          1..6646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1519..1537,1538..1543,1544..1560,1561..1566,
                     1567..1592)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1519..1537
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1602..1613
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4334..4353)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6206..6646,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga caattccgac
     1441 gtctaagaaa ccattattat catgacatta acctataaag ataggcgtat cacgaggccc
     1501 tttcgtcttc acctcgagtc cctatcagtg atagagattg acatccctat cagtgatagg
     1561 gatactgagc acatcagcag gacgcactga ccgaattcat taaagaggag aaaggtaccg
     1621 catgaaaata ataataaata atgatttccc ggtcgctaag gtcggagcgg atcaaattac
     1681 gactctagta agtgccaaag ttcatagttg catatatcgg ccaagattga gtatcgcgga
     1741 tggagccgct cccagagtat gcctttacag agccccacct ggatatggga aaaccgttgc
     1801 tcttgcgttc gagtggctac gccacagaac agccggacgt cctgcagtgt ggctttcttt
     1861 aagagccagt tcttacagtg aatttgatat ctgcgcagag attattgagc agcttgaaac
     1921 tttcgaaatg gtaaaattca gccgtgtgag agagggtgtg agcaagcctg cgctcttgcg
     1981 agaccttgca tctagtcttt ggcagagcac ctcgaataac gagatagaaa cgctagtttg
     2041 tttggataat attaatcatg acttagactt gccgttgttg cacgcactta tggagtttat
     2101 gttaaataca ccaaaaaata tcaggtttgc agttgcaggc aatacaataa aagggttctc
     2161 gcagcttaaa cttgcaggcg ctatgcggga gtacaccgag aaagacttgg cctttagcgc
     2221 agaagaggcg gtggcgttag cggaggcaga gtctgttctt ggagttcctg aagaacagat
     2281 agagaccttg gtgcaagaag ttgaggggtg gcctgctctt gtagtttttt tgttaaagcg
     2341 tgagttgccg gccaagcata tttcagcagt agttgaagta gacaattact ttagggatga
     2401 aatatttgag gcgattcccg agcgctatcg tgtttttctt gcaaattctt cattgctcga
     2461 tttcgtgacg cctgatcaat acaattatgt attcaaatgc gtcaatgggg tctcatgtat
     2521 taagtattta agcactaatt acatgttgct tcgccatgtg agcggtgagc cagcgcagtt
     2581 tacactgcat ccagtactgc gtaattttct acgagaaatt acttggactg aaaatcctgc
     2641 taaaagatcc tacctgctta agcgtgcagc tttctggcat tggcgtagag gtgaatacca
     2701 gtatgcaata cgaatatccc tacgggcgaa tgactgtcgc tgggcagtca gcatgtctga
     2761 gagaataatt ttagatttgt catttcgtca gggcgaaata gatgcgctga gacagtggct
     2821 gttagagctg ccgaagcagg cctggcacaa aaaacccata gtgcttatta gttacgcgtg
     2881 ggtattgtat ttcagtcagc aaggcgcgcg agcagagaag ttaattaaag acctatcttc
     2941 acaatccgat aaaaaaaata aatggcaaga aaaggaatgg ctgcagcttg tgcttgcaat
     3001 aggtaaagca acgaaagatg aaatgctttc gagtgaggag ctctgtaata agtggattag
     3061 tttatttggg gattcaaacg cagttggaaa aggggccgcg ctaacctgtt tggcttttat
     3121 ttttgccagt gagtatagat ttgcagagtt ggagaaggtg ctggctcagg cccaagccgt
     3181 gaataaattt gcaaaacaaa attttgcttt tggttggctg tatgtcgcga ggtttcaaca
     3241 agccctagca agcggaaaaa tgggctgggc gaggcagatt ataactcaag cacgcacaga
     3301 cagtcgcgcg cagatgatgg aatccgagtt tacttcgaaa atgtttgacg ctctagagct
     3361 tgagttacat tatgaattgc gctgcttgga cacctcagaa gaaaagctct ccaaaatttt
     3421 agagttcatt tccaatcacg gggtgacaga cgtgtttttt tccgtatgcc gtgctgtgtc
     3481 agcttggcgg cttggaagga gtgacctaaa tggctccatt gagatattgg agtgggcgaa
     3541 ggcgcatgcg gttgaaaaaa atctaccaag attggaagtt atgagccaaa ttgagatcta
     3601 tcagcgctta gtctgtcaag gcataacggg cataaataat ttaaaaactc ttgaagatca
     3661 taagattttc tccggacagc actcagcccc cctaaaagca cgcctgctgc ttgttcaatc
     3721 actagtgctt tcccgagatc ggaactttca tagtgccgcg cacagagcgt tattggctat
     3781 tcagcaagcc cgtaaaatta acgcgggcca gctggaagtc cgtggattat tgtgtttggc
     3841 cggagcgcag gcaggtgccg gtgatttaaa aaaggctcag cttaacattg tttatgcagt
     3901 ggagatagca aaacagcttc aatgctttca aacagttctt gatgaagtat gtttaattga
     3961 gcgaataata ccggcttcat gtgaagcctt cacagcagtt aatttagatc aagcgattgg
     4021 ggcttttagt cttccgcgaa tagttgagat tggaaagtcc gcagagaata aagctgacgc
     4081 tttattgaca cggaagcaga ttgctgtctt gaggcttgta aaagaggggt gctcaaacaa
     4141 acaaatagca acaaatatgc atgtcaccga agatgctata aagtggcata tgaggaaaat
     4201 atttgccacc ttgaatgtag tgaatcgcac gcaagcaaca attgaagctg agcgtcaagg
     4261 aattatctaa ggatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     4321 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     4381 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     4441 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     4501 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg accgatgccc
     4561 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     4621 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     4681 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     4741 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     4801 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     4861 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     4921 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     4981 ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcac tggaccgctg
     5041 atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta
     5101 ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
     5161 acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
     5221 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     5281 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct
     5341 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     5401 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     5461 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     5521 gtctggaaac gcggaagtcc cctacgtgct gctgaagttg cccgcaacag agagtggaac
     5581 caaccggtga taccacgata ctatgactga gagtcaacgc catgagcggc ctcatttctt
     5641 attctgagtt acaacagtcc gcaccgctgt ccggtagctc cttccggtgg gcgcggggca
     5701 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     5761 cggcagcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     5821 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
     5881 cacctacatc tgtattaacg aagcgctaac cgtttttatc aggctctggg aggcagaata
     5941 aatgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact ggcctcaggc
     6001 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     6061 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc
     6121 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcacca ggcgtttaag
     6181 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
     6241 gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga tgaacctgaa
     6301 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     6361 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     6421 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     6481 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     6541 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     6601 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacg
//
