LOCUS       sequence-434134-        5386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBR322 Origin, AmpR, with tdTER and Fdh genes under Vhb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5386)
  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 5386)
  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: ptrc99a-pVhb-tdTER-fdh
FEATURES             Location/Qualifiers
     source          1..5386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1478..1500
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2955..2972)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3125..3211
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3303..3330
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3349..3440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3441..3509,3510..4301)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3659..3678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4472..5060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4961..4980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5214..5231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5246..5386
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5312..5331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccccgacacc cgccaacacc cgctgacgcg ccctgacggt cacagcttgt ctgctcccgg
       61 catccgctta cagacaagct gtgaccgtct gacagcttat catcgactgc acgaaggttg
      121 aaaaataaaa acggcgctaa aaagcgccgt tttttttgac ggtggtagtg caccaatgct
      181 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtatta ttttttatca
      241 tgtttaccat acgctttggt cacatattcg ccgttcagca gaatgatatc ctgcggacga
      301 taatcaaatt tgccggtaaa aaagctttcc aggatatttt tggtgccttc cgcataacgg
      361 gtctgcgcat ctaaagtggt acccgagtaa tgcggtgtca tcgcattacc agcgccatat
      421 ttattacgca tatcgcgcca cggatgatct tttggagctg gttgcggaaa ccagacatcg
      481 ccgccgtaac cacgcagctg accgctttca agcgcggctg ccacatcttc cgccacgcaa
      541 atggcaccgc gcgcggtatt aaccagccag gcaccctttt taaacttgct caacagctct
      601 ttgttgatca ggcccttggt gcctgcatgc agcggcgcgt taacggtgac aatatctgcc
      661 tgggccacca gttcttcaat gttttcgaca cggcgcgcac ccactttctc ttcggcttct
      721 ttcggcagcg cctggtagtc atagtacagc agttctttcg gattgaacgg gagcaggcgt
      781 tccagtacgc gatagccaat gcgtcccgcg ccaatcgtgg caatggtttt accttcgata
      841 tcataagcat cttttgcgat cgccgccact tcccagtcat gattaatgat ttgctcatgg
      901 gcgggcacaa agttacgcac cagtaccagc atggtcatca ccacatgttc agcgacagac
      961 acgacgttgc tgcccgtcac ttccagcacg ctgatttttt tcccggtctg gttaatataa
     1021 tccagatcga tatggtcaga acccacgcca gccacaacca ctaatttcaa gtttttagct
     1081 ttgtccagac gttctttcgt gatataggcc ggatgaaacg gagtggttat gataatatca
     1141 gcgtccggaa tatgtttatc cagttcagag gtttcgcctt ccttatcgct tgtggtaatc
     1201 agttcatggc cctgatcctt cagccagttc gcaatgccca gcttgttttc ggtacaacca
     1261 tacagttttt cttcatccgc tgcgtgtttg cccgcatcat acaaaaccag gacaattttc
     1321 atgtgtgcct ccctaactac ttaattcata agttcattat tatggttttg ccatacatca
     1381 gtatggtgta gcagcactta ttataatctt tattgcctct taaaacttaa tccacatcaa
     1441 aactcaaata cttttaaccc cagcgtcctg taagcttccg ggagctgcat gtgtcagagg
     1501 ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag
     1561 cggcatgcat ttacgttaag cttacaggac gctggggtta aaagtatttg agttttgatg
     1621 tggattaagt tttaagaggc aataaagatt ataataagtg ctgctacacc atactgatgt
     1681 atggcaaaac cataataatg aacttaagac gacaaggggg cgaatttaat gattgtgaag
     1741 ccgatggtac gcaacaacat ttgtctgaac gcgcacccgc agggatgcaa gaaaggcgtc
     1801 gaagatcaga ttgaatatac caagaagcgt atcactgcag aggtgaaggc cggggctaaa
     1861 gcccctaaaa atgtcctggt gctcggctgt agcaacggtt acggtctggc aagtcgcatc
     1921 actgcggctt tcggttacgg ggccgctacc attggcgtct cgtttgaaaa ggcaggtagc
     1981 gaaactaaat atggcactcc gggttggtat aataacctgg catttgacga agccgcgaaa
     2041 cgcgaaggtt tatattcagt gacgattgat ggcgacgcgt tcagcgacga aatcaaagcg
     2101 caggtgatcg aggaagcaaa gaagaaaggt atcaaatttg atctgatcgt gtactcgctg
     2161 gcatcgccgg ttcgcaccga cccggatacc ggaattatgc acaaatctgt gctgaagccg
     2221 tttggtaaaa cctttacagg taaaaccgtg gatccgttta ctggagagct taaagagatc
     2281 tcggcagaac ccgcgaatga tgaggaagca gcggcgaccg tgaaagtgat gggaggcgaa
     2341 gattgggaac gctggatcaa acaactgtcg aaagaaggcc tgttagaaga gggctgcatt
     2401 acacttgcgt atagttacat cggtccggaa gcgacccaag cactctatcg taagggtact
     2461 atcggtaaag cgaaagaaca ccttgaagcg actgcacacc gcctgaacaa agaaaacccg
     2521 tctatccgtg cgttcgtcag cgtaaacaaa ggtttggtta cgcgtgcgag cgcggttatc
     2581 cctgttattc cgctgtattt ggccagtttg tttaaagtta tgaaggaaaa aggcaaccat
     2641 gaagggtgca ttgaacagat tacacgcctt tacgccgagc gcctgtatcg taaagatgga
     2701 acgatccctg tggatgaaga aaaccgcatt cgcatcgatg attgggaatt ggaagaggat
     2761 gtgcagaagg ccgtttcggc cctgatggaa aaagtgaccg gcgaaaacgc cgaatcttta
     2821 actgatctcg ccgggtaccg ccatgatttt ctggcaagca acggtttcga tgtggagggt
     2881 attaactatg aggcggaggt tgaacgcttc gatcgcattt aaggctgttt tggcggatga
     2941 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     3001 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     3061 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     3121 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3181 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     3241 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     3301 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     3361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3421 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3481 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3541 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3601 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3661 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3721 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3781 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3841 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3901 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3961 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4021 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4081 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4141 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4201 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4261 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5161 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5221 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5281 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5341 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcg
//
