LOCUS       sequence-60520-e        6633 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 6633)
  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 6633)
  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-alkSm4-3
FEATURES             Location/Qualifiers
     source          1..6633
                     /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"
     primer_bind     1467..1485
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1506..1524,1525..1530,1531..1547,1548..1553,
                     1554..1579)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1506..1524
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1531..1549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1589..1600
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4321..4340)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6193..6633,1..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 taagaaacca
     1441 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcacc
     1501 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
     1561 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccgcat gaaaataata
     1621 ataaataatg atttcccggt cgctaaggtc ggagcggatc aaattacgac tctagtaagt
     1681 gccaaagttc atagttgcat atatcggcca agattgagta tcgcggatgg agccgctccc
     1741 agagtatgcc tttacagagc cccacctgga tatgggaaaa ccgttgctct tgcgttcgag
     1801 tggctacgcc acagaacagc cggacgtcct gcagtgtggc tttctttaag agccagttct
     1861 tacagtgaat ttgatatctg cgcagagatt attgagcagc ttgaaacttt cgaaatggta
     1921 aaattcagcc gtgtgagaga gggtgtgagc aagcctgcgc tcttgcgaga ccttgcatct
     1981 agtctttggc agagcacctc gaataacgag atagaaacgc tagtttgttt ggataatatt
     2041 aatcatgact tagacttgcc gttgttgcac gcacttatgg agtttatgtt aaatacacca
     2101 aaaaatatca ggtttgcagt tgcaggcaat acaataaaag ggttctcgca gcttaaactt
     2161 gcaggcgcta tgcgggagta caccgagaaa gacttggcct ttagcgcaga agaggcggtg
     2221 gcgttagcgg aggcagagtc tgttcttgga gttcctgaag aacagataga gaccttggtg
     2281 caagaagttg aggggtggcc tgctcttgta gtttttttgt taaagcgtga gttgccggcc
     2341 aagcatattt cagcagtagt tgaagtagac aattacttta gggatgaaat atttgaggcg
     2401 attcccgagc gctatcgtgt ttttcttgca aattcttcat tgctcgattt cgtgacgcct
     2461 gatcaataca attatgtatt caaatgcgtc aatggggtct catgtattaa gtatttaagc
     2521 actaattaca tgttgcttcg ccatgtgagc ggtgagccag cgcagtttac actgcatcca
     2581 gtactgcgta attttctacg agaaattact tggactgaaa atcctgctaa aagatcctac
     2641 ctgcttaagc gtgcagcttt ctggcattgg cgtagaggtg aataccagta tgcaatacga
     2701 atatccctac gggcgaatga ctgtcgctgg gcagtcagca tgtctgagag aataatttta
     2761 gatttgtcat ttcgtcaggg cgaaatagat gcgctgagac agtggctgtt agagctgccg
     2821 aagcaggcct ggcacataaa acccatagtg cttattagtt acgcgtgggt attgtatttc
     2881 agtcagcaag gcgcgcgagc agagaagtta attaaagacc tatcttcaca atccgataaa
     2941 aaaaataaat ggcaagaaaa ggaatggctg cagcttgtgc ttgcaatagg taaagcaacg
     3001 aaagatgaaa tgcttacgag tgaggagctc tgtaataagt ggattagttt atttggggat
     3061 tcaaacgcag ttggaaaagg ggccgcgcta acctgtttgg cttttatttt tgccagtgag
     3121 tatagatttg cagagttgga gaaggtgctg gctcaggccc aagccgtgaa taaatttgca
     3181 aaacaaaatt ttgcttttgg ttggctgtat gtcgcgaggt ttcaacaagc cctagcaagc
     3241 ggagaaatgg gctgggcgag gcagattata actcaagcac gcacagacag tcgcgcgcag
     3301 atgatggaat ccgagtttac ttcgaaaatg tttgacgctc tagagcttga gttacattat
     3361 gaattgcgct gcttggacac ctcagaagaa aagctctcca aaattttaga gttcatttcc
     3421 aatcacgggg tgacagacgt gtttttttcc gtatgccgtg ctgtgtcagc ttggcggctt
     3481 ggaaggagtg acctaaatgg ctccattgag atattggagt gggcgaaggc gcatgcagtt
     3541 gaaaaaaatc taccaagatt ggaagttatg agccaaattg agatctatca gcgcttagtc
     3601 tgtcaaggca taacgggcat aaataattta aaaactcttg aagatcataa gattttctcc
     3661 ggacagcact cagcccccct aaaagcacgc ctgctgcttg ttcaatcact agtgctttcc
     3721 cgagatcgga actttcatag tgccgcgcac agagcgttat tggctattca gcaagcccgt
     3781 aaaattaacg cgggccagct ggaagtccgt ggattattgt gtttggccgg agcgcaggca
     3841 ggtgccggtg atttaaaaaa ggctcagctt aacattgttt atgcagtgga gatagcaaaa
     3901 cagcttcaat gctttcaaac agttcttgat gatgtatgtt taattgagcg aataattccg
     3961 gcttcatgtg aagccttcac agcagttaat ttagaccaag cgattggggc ttttagtctt
     4021 ccgcgaatag ttgagattgg aaagtccgca gagaataaag ctgacgcttt attgacacgg
     4081 aagcagattg ctgtcttgag gcttgtaaaa gaggggtgct caaacaaaca aatagcaaca
     4141 aatatgcatg tcaccgaaga tgctataaag tggcatatga ggaaaatatt tgccaccttg
     4201 aatgtagtga atcgcacgca agcaacaatt gaagctgagc gtcaaggaat tatctaagga
     4261 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     4321 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     4381 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     4441 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     4501 gcttcctaat gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca
     4561 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     4621 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     4681 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     4741 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     4801 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     4861 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     4921 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     4981 gatcgctcgc ggctcttacc agcctaactt cgatcactgg accgctgatc gtcacggcga
     5041 tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat
     5101 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
     5161 tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt
     5221 cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc
     5281 catctccagc agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg
     5341 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     5401 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     5461 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     5521 gaagtcccct acgtgctgct gaagttgccc gcaacagaga gtggaaccaa ccggtgatac
     5581 cacgatacta tgactgagag tcaacgccat gagcggcctc atttcttatt ctgagttaca
     5641 acagtccgca ccgctgtccg gtagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     5701 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgcccaa
     5761 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc cctgcaccat
     5821 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     5881 attaacgaag cgctaaccgt ttttatcagg ctctgggagg cagaataaat gatcatatcg
     5941 tcaattatta cctccacggg gagagcctga gcaaactggc ctcaggcatt tgagaagcac
     6001 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     6061 atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat
     6121 tcatccgctt attatcactt attcaggcgt agcaccaggc gtttaagggc accaataact
     6181 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     6241 cattctgccg acatggaagc catcacagac ggcatgatga acctgaatcg ccagcggcat
     6301 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     6361 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     6421 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     6481 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     6541 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     6601 tatcaccagc tcaccgtctt tcattgccat acg
//
