LOCUS       sequence-8740-ex        8759 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 8759)
  AUTHORS   Pitera DJ, Paddon CJ, Newman JD, Keasling JD
  TITLE     Balancing a heterologous mevalonate pathway for improved isoprenoid 
            production in Escherichia coli.
  JOURNAL   Metab Eng. 2007 Mar . 9(2):193-207.
  PUBMED    17239639
REFERENCE   2  (bases 1 to 8759)
  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. 
            2007 Mar . 9(2):193-207."
COMMENT     Sequence Label: pBAD24MevT
FEATURES             Location/Qualifiers
     source          1..8759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(2063..2074)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4960..4971
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5616..5633)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5786..5872
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5964..5991
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6010..6101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6102..6170,6171..6962)
                     /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(6320..6339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7004..7459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7091..7110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      7570..8158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8059..8078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8312..8329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8495..8517
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8656..8675
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tggtacccgg ggatcctcta gagtcgacta ggaggaatat aaaatgaaaa attgtgtcat
     1381 cgtcagtgcg gtacgtactg ctatcggtag ttttaacggt tcactcgctt ccaccagcgc
     1441 catcgacctg ggggcgacag taattaaagc cgccattgaa cgtgcaaaaa tcgattcaca
     1501 acacgttgat gaagtgatta tgggtaacgt gttacaagcc gggctggggc aaaatccggc
     1561 gcgtcaggca ctgttaaaaa gcgggctggc agaaacggtg tgcggattca cggtcaataa
     1621 agtatgtggt tcgggtctta aaagtgtggc gcttgccgcc caggccattc aggcaggtca
     1681 ggcgcagagc attgtggcgg ggggtatgga aaatatgagt ttagccccct acttactcga
     1741 tgcaaaagca cgctctggtt atcgtcttgg agacggacag gtttatgacg taatcctgcg
     1801 cgatggcctg atgtgcgcca cccatggtta tcatatgggg attaccgccg aaaacgtggc
     1861 taaagagtac ggaattaccc gtgaaatgca ggatgaactg gcgctacatt cacagcgtaa
     1921 agcggcagcc gcaattgagt ccggtgcttt tacagccgaa atcgtcccgg taaatgttgt
     1981 cactcgaaag aaaaccttcg tcttcagtca agacgaattc ccgaaagcga attcaacggc
     2041 tgaagcgtta ggtgcattgc gcccggcctt cgataaagca ggaacagtca ccgctgggaa
     2101 cgcgtctggt attaacgacg gtgctgccgc tctggtgatt atggaagaat ctgcggcgct
     2161 ggcagcaggc cttacccccc tggctcgcat taaaagttat gccagcggtg gcgtgccccc
     2221 cgcattgatg ggtatggggc cagtacctgc cacgcaaaaa gcgttacaac tggcggggct
     2281 gcaactggcg gatattgatc tcattgaggc taatgaagca tttgctgcac agttccttgc
     2341 cgttgggaaa aacctgggct ttgattctga gaaagtgaat gtcaacggcg gggccatcgc
     2401 gctcgggcat cctatcggtg ccagtggtgc tcgtattctg gtcacactat tacatgccat
     2461 gcaggcacgc gataaaacgc tggggctggc aacactgtgc attggcggcg gtcagggaat
     2521 tgcgatggtg attgaacggt tgaattaagg aggacagcta aatgaaactc tcaactaaac
     2581 tttgttggtg tggtattaaa ggaagactta ggccgcaaaa gcaacaacaa ttacacaata
     2641 caaacttgca aatgactgaa ctaaaaaaac aaaagaccgc tgaacaaaaa accagacctc
     2701 aaaatgtcgg tattaaaggt atccaaattt acatcccaac tcaatgtgtc aaccaatctg
     2761 agctagagaa atttgatggc gtttctcaag gtaaatacac aattggtctg ggccaaacca
     2821 acatgtcttt tgtcaatgac agagaagata tctactcgat gtccctaact gttttgtcta
     2881 agttgatcaa gagttacaac atcgacacca acaaaattgg tagattagaa gtcggtactg
     2941 aaactctgat tgacaagtcc aagtctgtca agtctgtctt gatgcaattg tttggtgaaa
     3001 acactgacgt cgaaggtatt gacacgctta atgcctgtta cggtggtacc aacgcgttgt
     3061 tcaactcttt gaactggatt gaatctaacg catgggatgg tagagacgcc attgtagttt
     3121 gcggtgatat tgccatctac gataagggtg ccgcaagacc aaccggtggt gccggtactg
     3181 ttgctatgtg gatcggtcct gatgctccaa ttgtatttga ctctgtaaga gcttcttaca
     3241 tggaacacgc ctacgatttt tacaagccag atttcaccag cgaatatcct tacgtcgatg
     3301 gtcatttttc attaacttgt tacgtcaagg ctcttgatca agtttacaag agttattcca
     3361 agaaggctat ttctaaaggg ttggttagcg atcccgctgg ttcggatgct ttgaacgttt
     3421 tgaaatattt cgactacaac gttttccatg ttccaacctg taaattggtc acaaaatcat
     3481 acggtagatt actatataac gatttcagag ccaatcctca attgttccca gaagttgacg
     3541 ccgaattagc tactcgcgat tatgacgaat ctttaaccga taagaacatt gaaaaaactt
     3601 ttgttaatgt tgctaagcca ttccacaaag agagagttgc ccaatctttg attgttccaa
     3661 caaacacagg taacatgtac accgcatctg tttatgccgc ctttgcatct ctattaaact
     3721 atgttggatc tgacgactta caaggcaagc gtgttggttt attttcttac ggttccggtt
     3781 tagctgcatc tctatattct tgcaaaattg ttggtgacgt ccaacatatt atcaaggaat
     3841 tagatattac taacaaatta gccaagagaa tcaccgaaac tccaaaggat tacgaagctg
     3901 ccatcgaatt gagagaaaat gcccatttga agaagaactt caaacctcaa ggttccattg
     3961 agcatttgca aagtggtgtt tactacttga ccaacatcga tgacaaattt agaagatctt
     4021 acgatgttaa aaaataagga ggattacact atggttttaa ccaataaaac agtcatttct
     4081 ggatcgaaag tcaaaagttt atcatctgcg caatcgagct catcaggacc ttcatcatct
     4141 agtgaggaag atgattcccg cgatattgaa agcttggata agaaaatacg tcctttagaa
     4201 gaattagaag cattattaag tagtggaaat acaaaacaat tgaagaacaa agaggtcgct
     4261 gccttggtta ttcacggtaa gttacctttg tacgctttgg agaaaaaatt aggtgatact
     4321 acgagagcgg ttgcggtacg taggaaggct ctttcaattt tggcagaagc tcctgtatta
     4381 gcatctgatc gtttaccata taaaaattat gactacgacc gcgtatttgg cgcttgttgt
     4441 gaaaatgtta taggttacat gcctttgccc gttggtgtta taggcccctt ggttatcgat
     4501 ggtacatctt atcatatacc aatggcaact acagagggtt gtttggtagc ttctgccatg
     4561 cgtggctgta aggcaatcaa tgctggcggt ggtgcaacaa ctgttttaac taaggatggt
     4621 atgacaagag gcccagtagt ccgtttccca actttgaaaa gatctggtgc ctgtaagata
     4681 tggttagact cagaagaggg acaaaacgca attaaaaaag cttttaactc tacatcaaga
     4741 tttgcacgtc tgcaacatat tcaaacttgt ctagcaggag atttactctt catgagattt
     4801 agaacaacta ctggtgacgc aatgggtatg aatatgattt ctaaaggtgt cgaatactca
     4861 ttaaagcaaa tggtagaaga gtatggctgg gaagatatgg aggttgtctc cgtttctggt
     4921 aactactgta ccgacaaaaa accagctgcc atcaactgga tcgaaggtcg tggtaagagt
     4981 gtcgtcgcag aagctactat tcctggtgat gttgtcagaa aagtgttaaa aagtgatgtt
     5041 tccgcattgg ttgagttgaa cattgctaag aatttggttg gatctgcaat ggctgggtct
     5101 gttggtggat ttaacgcaca tgcagctaat ttagtgacag ctgttttctt ggcattagga
     5161 caagatcctg cacaaaatgt tgaaagttcc aactgtataa cattgatgaa agaagtggac
     5221 ggtgatttga gaatttccgt atccatgcca tccatcgaag taggtaccat cggtggtggt
     5281 actgttctag aaccacaagg tgccatgttg gacttattag gtgtaagagg cccgcatgct
     5341 accgctcctg gtaccaacgc acgtcaatta gcaagaatag ttgcctgtgc cgtcttggca
     5401 ggtgaattat ccttatgtgc tgccctagca gccggccatt tggttcaaag tcatatgacc
     5461 cacaacagga aacctgctga accaacaaaa cctaacaatt tggacgccac tgatataaat
     5521 cgtttgaaag atgggtccgt cacctgcatt aaatcctaag tcgacctgca ggcatgcaag
     5581 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     5641 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     5701 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     5761 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     5821 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     5881 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     5941 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     6001 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6061 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6121 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6181 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6241 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6301 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     6361 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6421 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6481 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6541 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6601 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6661 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6721 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6781 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6841 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6901 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6961 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     7021 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     7081 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     7141 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     7201 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     7261 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     7321 tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     7381 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     7441 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7501 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7561 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7621 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7681 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     7741 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7801 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7861 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7921 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7981 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8041 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8101 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8161 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     8221 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     8281 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     8341 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggtc
     8401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     8461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     8521 tcaccgtcat caccgaaacg cgcgaggcag caaggagatg gcgcccaaca gtcccccggc
     8581 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     8641 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     8701 ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatc
//
