LOCUS       sequence-431943-        7670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pACYC Origin, CmR, with vfFadBA gene under Vhb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7670)
  AUTHORS   Mehrer CR, Incha MR, Politz MC, Pfleger BF
  TITLE     Anaerobic production of medium-chain fatty alcohols via a 
            beta-reduction pathway.
  JOURNAL   Metab Eng. 2018 Jul;48:63-71. doi: 10.1016/j.ymben.2018.05.011. Epub
            2018 May 25.
  PUBMED    29807110
REFERENCE   2  (bases 1 to 7670)
  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: "Metab 
            Eng."; date: "2018-07"; volume: "48:63-71. doi"; pages: " 
            10.1016/j.ymben.2018.05.011. Epub 2018 May 25"
COMMENT     Sequence Label: pACYC-pVhb-vfFadBA
FEATURES             Location/Qualifiers
     source          1..7670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             272..280
                     /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)"
     misc_feature    3642..3698
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3731..3748)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3901..3987
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4079..4106
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4125..4216
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4674..5129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4761..4780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4971..4992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5685..6344)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6259..6278
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6345..6447)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6839..6856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6973..7518)
                     /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(7053..7072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7594..7613
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggatctgctc atgtttgaca gcttatcatc gatgcataat gtgcctgtca aatggacgaa
       61 gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc
      121 aactcgagaa gcttacagga cgctggggtt aaaagtattt gagttttgat gtggattaag
      181 ttttaagagg caataaagat tataataagt gctgctacac catactgatg tatggcaaaa
      241 ccataataat gaacttagca catcaccaat ctaaggaggt aagaaatgat ttatcaaggt
      301 gaaaacctct ctgtagacta catcgaaccg ggcattgctc acttagtatt taatgctaaa
      361 ggcagtgtaa ataagctcaa cttagccacc ctacaatcgg ttggtgaagc catagatgca
      421 ctctactcac aaaaggactt acaaggtcta cttctatcat cagataaaag tgcatttatt
      481 gttggtgctg atattacgga gtttcttggt ttgtttgata ttccaacaga agaactcagt
      541 gattggttac atcaagctaa cgccattttt aatcgtctag aagacattcc agtacctact
      601 ttatctgcta ttactggatt tgctttagga ggcggttgtg aatgtgtcct tgctacggat
      661 tttcgattag cagatgaaac ggcatctatc ggtctacccg aaacacagct tgggatcatg
      721 cccggctggg gtggctcagt gcgcttacct cgcttaattg gtgccgatcc agcaatggaa
      781 gtcatcacaa cgggtaagcc aaagcgagca aaagatgcct tagctctggg aatggttgat
      841 ggtgttgttt cacgtgaaac cttgataaca gattctgtca ctatgcttaa aaaagcgata
      901 gagggtcaac tggattggca atctcgtcgt acacagaaaa aagcaccatt aaaactctct
      961 cctctagagg cagcaatgtc atttaatgtc gcaaaaggca tgataatgaa aatggcaggt
     1021 aaacattacc ctgcaccaat gacagcagta aagtccattg aaaatagtgc cttcatggat
     1081 cgtgatgctg ctcttgaagt tgaaaataaa aacttcgttg cactcactca aacagatgtg
     1141 gctaagtcat tagttggtat tttcttaaac gatcaattag tgaagagtaa agcgaaacaa
     1201 gccataaaaa atagtgaacc cgtgactagc gcagcggttc ttggtgctgg aattatgggt
     1261 ggaggtattg cttatcaatc cgcatcaaaa ggcgtccctg tattaatgaa ggacattgct
     1321 caacaatcat tagatttagg catgagtgaa gcctctaaac tgcttaataa acaattagaa
     1381 cgaggacgtt tatctggttt aaagatggca aaagtactct cttctattac cccatcacta
     1441 aactatgcca gtattgaaaa caaagatatt gtcgtagaag ccgtcgttga gaacccaaag
     1501 gttaaagctg cagtattagc tgaagttgaa aatgaagtga atgagcacgc catcttagcc
     1561 tcaaatacat cgacaatacc tatttcctta ctagcaaaat cattaaaacg cccagaaaat
     1621 ttctgtggca tgcacttctt caatcctgta catcggatgc cactcgttga agtgatccgt
     1681 ggtgaaaaaa cgtcacaaca aaccattgat cgtgttgttg cttacgcttc tcaaatggga
     1741 aaaacaccta ttgtcatcaa tgactgccct ggattttttg ttaaccgagt tcttttccct
     1801 tatttcgcag ggttcagttt gttgctgcgt gatggtggtg actatcaaca aatagataac
     1861 gtcatggaaa aagagtttgg ttggccaatg ggaccagctt atttacttga tgtcgttgga
     1921 attgatacag cacatcacgc acaagccgtc atggcacaag gcttccctga gcgaatggca
     1981 aaaaatggac gagatgtgat tgatgcgatg ttcgaatccg atcgctatgg tcaaaaaaat
     2041 ggaagtggtt tttatgcata tagcatagat agaaaaggca aacctaaaaa gaacattgat
     2101 gaaagcacag caggcattat tgcgacagta accaatacaa cgcaacctta tacatcagag
     2161 cagatcagtg ctcgcatgat gatacctatg attaatgagg tcatccgctg cctagatgaa
     2221 ggaattattg cctctcctgc agaagctgac atggcattgg tttatggttt gggcttccct
     2281 ccatttaaag gaggagtatt ccgctatctt gatgcaattg gattaaccac ttatctcgac
     2341 atggcaaaag aatttgaaca tctaggtgcc gtttatcaag taccagaaag catcaaacaa
     2401 aaagcagccg atgatgaatg ttattaccca gcgcctcaat cattatcagc accgtcagcc
     2461 tagggaagga ttaatcatga aaaacgtagt tattgttgac tgtattcgaa cacctatggg
     2521 tcgctcaaaa aacggtgtat ttcgtcatac tcgcgctgaa gatttatcag cgcacctaat
     2581 gaaaggtttg ctaaaacgaa atccaagtgt cgatcctaat gacattgaag atatctattg
     2641 gggatgtgta caacaaacgt tagaacaagg cttcaatatt gcacgtaatt ccgcactact
     2701 tgccggttta ccacaaagta ttgccgcaac aactgttaac cgtttatgtg gctcatccat
     2761 gcaagctctg catgatgcat cacgtgccat tatggttggc gatgccgaaa tttgcataat
     2821 tggtggtgtt gaacacatgg ggcatgtacc aatgaaccat ggcgttgatt tccattcagg
     2881 actttcaaaa agtgttgcta aagcatcagg aatgatggga ttaacagctg aaatgctagg
     2941 taaaatgcac ggtatatctc gagagcagca agatgccttt gcgctcgcat cccatcaaaa
     3001 agcacataaa gcaacgattg aaggctattt tgattcggag atccttccaa tggaaggtca
     3061 cgatgaaaat ggcgctttaa ccttagtcac acatgatgag gtcattcgac ctgaaacaac
     3121 attagaagga ttagccgcat tacgaccagc tttcgatcct gcaaatggta cagtaacagc
     3181 aggcagttca tcggcattat cggatggcgc atcagcgatg ctagttatga gcgaagagaa
     3241 agccaatgag ttaggtttac ctattcgagc gaaagttcgc tcaatggcgg tatctggttg
     3301 cgatccatct attatgggat acggccctgt tccagcgacg aaaaaagcat taaaacgtgc
     3361 cggattatca ctggatgata tagagctatt cgaactgaat gaagcgtttg ccgctcaatc
     3421 actaccttgt attaaagatt taggcttact tgatgtgatg gatgaaaagg tcaacttaaa
     3481 tggtggtgct atcgctcttg gtcatcctct gggttgctct ggatctcgca ttgccaccac
     3541 attaatcaat aatatggagc gaactggagc caagttaggt gttgcaacaa tgtgtattgg
     3601 attagggcaa gggattgcaa ctgtgtttga gcgcccttaa tgaattcgag ctcggtaccc
     3661 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc ggatgagaga
     3721 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
     3781 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
     3841 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
     3901 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     3961 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     4021 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     4081 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
     4141 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4201 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4261 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4321 agatcagttg ggtgcagcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4381 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4441 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4501 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4561 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4621 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc
     4681 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4741 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4801 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4861 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4921 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4981 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     5041 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5101 ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata
     5161 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5221 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5281 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5341 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5401 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5461 tcctgttacc agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     5521 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     5581 cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg
     5641 tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     5701 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     5761 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     5821 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     5881 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     5941 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     6001 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     6061 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     6121 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     6181 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     6241 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     6301 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     6361 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     6421 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     6481 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     6541 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     6601 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     6661 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
     6721 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     6781 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     6841 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     6901 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     6961 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     7021 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     7081 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     7141 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     7201 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     7261 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     7321 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     7381 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     7441 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     7501 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctaggctcat
     7561 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     7621 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
//
