LOCUS       sequence-8737-ex        9573 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 9573)
  AUTHORS   Martin VJ, Pitera DJ, Withers ST, Newman JD, Keasling JD
  TITLE     Engineering a mevalonate pathway in Escherichia coli for production 
            of terpenoids.
  JOURNAL   Nat Biotechnol. 2003 Jul . 21(7):796-802.
  PUBMED    12778056
REFERENCE   2  (bases 1 to 9573)
  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: "Nat 
            Biotechnol. 2003 Jul . 21(7):796-802."
COMMENT     Sequence Label: pBAD33MevT
FEATURES             Location/Qualifiers
     source          1..9573
                     /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(2059..2070)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4956..4967
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5612..5629)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5782..5868
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5960..5987
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6006..6097
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6555..7010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6642..6661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(7565..8224)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8139..8158
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(8225..8327)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(8719..8736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8853..9398)
                     /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(8933..8952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9470..9489
                     /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 ctagcgaatt cgagctcggt
     1321 acccggggat cctctagagt cgactaggag gaatataaaa tgaaaaattg tgtcatcgtc
     1381 agtgcggtac gtactgctat cggtagtttt aacggttcac tcgcttccac cagcgccatc
     1441 gacctggggg cgacagtaat taaagccgcc attgaacgtg caaaaatcga ttcacaacac
     1501 gttgatgaag tgattatggg taacgtgtta caagccgggc tggggcaaaa tccggcgcgt
     1561 caggcactgt taaaaagcgg gctggcagaa acggtgtgcg gattcacggt caataaagta
     1621 tgtggttcgg gtcttaaaag tgtggcgctt gccgcccagg ccattcaggc aggtcaggcg
     1681 cagagcattg tggcgggggg tatggaaaat atgagtttag ccccctactt actcgatgca
     1741 aaagcacgct ctggttatcg tcttggagac ggacaggttt atgacgtaat cctgcgcgat
     1801 ggcctgatgt gcgccaccca tggttatcat atggggatta ccgccgaaaa cgtggctaaa
     1861 gagtacggaa ttacccgtga aatgcaggat gaactggcgc tacattcaca gcgtaaagcg
     1921 gcagccgcaa ttgagtccgg tgcttttaca gccgaaatcg tcccggtaaa tgttgtcact
     1981 cgaaagaaaa ccttcgtctt cagtcaagac gaattcccga aagcgaattc aacggctgaa
     2041 gcgttaggtg cattgcgccc ggccttcgat aaagcaggaa cagtcaccgc tgggaacgcg
     2101 tctggtatta acgacggtgc tgccgctctg gtgattatgg aagaatctgc ggcgctggca
     2161 gcaggcctta cccccctggc tcgcattaaa agttatgcca gcggtggcgt gccccccgca
     2221 ttgatgggta tggggccagt acctgccacg caaaaagcgt tacaactggc ggggctgcaa
     2281 ctggcggata ttgatctcat tgaggctaat gaagcatttg ctgcacagtt ccttgccgtt
     2341 gggaaaaacc tgggctttga ttctgagaaa gtgaatgtca acggcggggc catcgcgctc
     2401 gggcatccta tcggtgccag tggtgctcgt attctggtca cactattaca tgccatgcag
     2461 gcacgcgata aaacgctggg gctggcaaca ctgtgcattg gcggcggtca gggaattgcg
     2521 atggtgattg aacggttgaa ttaaggagga cagctaaatg aaactctcaa ctaaactttg
     2581 ttggtgtggt attaaaggaa gacttaggcc gcaaaagcaa caacaattac acaatacaaa
     2641 cttgcaaatg actgaactaa aaaaacaaaa gaccgctgaa caaaaaacca gacctcaaaa
     2701 tgtcggtatt aaaggtatcc aaatttacat cccaactcaa tgtgtcaacc aatctgagct
     2761 agagaaattt gatggcgttt ctcaaggtaa atacacaatt ggtctgggcc aaaccaacat
     2821 gtcttttgtc aatgacagag aagatatcta ctcgatgtcc ctaactgttt tgtctaagtt
     2881 gatcaagagt tacaacatcg acaccaacaa aattggtaga ttagaagtcg gtactgaaac
     2941 tctgattgac aagtccaagt ctgtcaagtc tgtcttgatg caattgtttg gtgaaaacac
     3001 tgacgtcgaa ggtattgaca cgcttaatgc ctgttacggt ggtaccaacg cgttgttcaa
     3061 ctctttgaac tggattgaat ctaacgcatg ggatggtaga gacgccattg tagtttgcgg
     3121 tgatattgcc atctacgata agggtgccgc aagaccaacc ggtggtgccg gtactgttgc
     3181 tatgtggatc ggtcctgatg ctccaattgt atttgactct gtaagagctt cttacatgga
     3241 acacgcctac gatttttaca agccagattt caccagcgaa tatccttacg tcgatggtca
     3301 tttttcatta acttgttacg tcaaggctct tgatcaagtt tacaagagtt attccaagaa
     3361 ggctatttct aaagggttgg ttagcgatcc cgctggttcg gatgctttga acgttttgaa
     3421 atatttcgac tacaacgttt tccatgttcc aacctgtaaa ttggtcacaa aatcatacgg
     3481 tagattacta tataacgatt tcagagccaa tcctcaattg ttcccagaag ttgacgccga
     3541 attagctact cgcgattatg acgaatcttt aaccgataag aacattgaaa aaacttttgt
     3601 taatgttgct aagccattcc acaaagagag agttgcccaa tctttgattg ttccaacaaa
     3661 cacaggtaac atgtacaccg catctgttta tgccgccttt gcatctctat taaactatgt
     3721 tggatctgac gacttacaag gcaagcgtgt tggtttattt tcttacggtt ccggtttagc
     3781 tgcatctcta tattcttgca aaattgttgg tgacgtccaa catattatca aggaattaga
     3841 tattactaac aaattagcca agagaatcac cgaaactcca aaggattacg aagctgccat
     3901 cgaattgaga gaaaatgccc atttgaagaa gaacttcaaa cctcaaggtt ccattgagca
     3961 tttgcaaagt ggtgtttact acttgaccaa catcgatgac aaatttagaa gatcttacga
     4021 tgttaaaaaa taaggaggat tacactatgg ttttaaccaa taaaacagtc atttctggat
     4081 cgaaagtcaa aagtttatca tctgcgcaat cgagctcatc aggaccttca tcatctagtg
     4141 aggaagatga ttcccgcgat attgaaagct tggataagaa aatacgtcct ttagaagaat
     4201 tagaagcatt attaagtagt ggaaatacaa aacaattgaa gaacaaagag gtcgctgcct
     4261 tggttattca cggtaagtta cctttgtacg ctttggagaa aaaattaggt gatactacga
     4321 gagcggttgc ggtacgtagg aaggctcttt caattttggc agaagctcct gtattagcat
     4381 ctgatcgttt accatataaa aattatgact acgaccgcgt atttggcgct tgttgtgaaa
     4441 atgttatagg ttacatgcct ttgcccgttg gtgttatagg ccccttggtt atcgatggta
     4501 catcttatca tataccaatg gcaactacag agggttgttt ggtagcttct gccatgcgtg
     4561 gctgtaaggc aatcaatgct ggcggtggtg caacaactgt tttaactaag gatggtatga
     4621 caagaggccc agtagtccgt ttcccaactt tgaaaagatc tggtgcctgt aagatatggt
     4681 tagactcaga agagggacaa aacgcaatta aaaaagcttt taactctaca tcaagatttg
     4741 cacgtctgca acatattcaa acttgtctag caggagattt actcttcatg agatttagaa
     4801 caactactgg tgacgcaatg ggtatgaata tgatttctaa aggtgtcgaa tactcattaa
     4861 agcaaatggt agaagagtat ggctgggaag atatggaggt tgtctccgtt tctggtaact
     4921 actgtaccga caaaaaacca gctgccatca actggatcga aggtcgtggt aagagtgtcg
     4981 tcgcagaagc tactattcct ggtgatgttg tcagaaaagt gttaaaaagt gatgtttccg
     5041 cattggttga gttgaacatt gctaagaatt tggttggatc tgcaatggct gggtctgttg
     5101 gtggatttaa cgcacatgca gctaatttag tgacagctgt tttcttggca ttaggacaag
     5161 atcctgcaca aaatgttgaa agttccaact gtataacatt gatgaaagaa gtggacggtg
     5221 atttgagaat ttccgtatcc atgccatcca tcgaagtagg taccatcggt ggtggtactg
     5281 ttctagaacc acaaggtgcc atgttggact tattaggtgt aagaggcccg catgctaccg
     5341 ctcctggtac caacgcacgt caattagcaa gaatagttgc ctgtgccgtc ttggcaggtg
     5401 aattatcctt atgtgctgcc ctagcagccg gccatttggt tcaaagtcat atgacccaca
     5461 acaggaaacc tgctgaacca acaaaaccta acaatttgga cgccactgat ataaatcgtt
     5521 tgaaagatgg gtccgtcacc tgcattaaat cctaagtcga cctgcaggca tgcaagcttg
     5581 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     5641 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     5701 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     5761 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     5821 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     5881 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     5941 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     6001 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6181 ggtgaaagta aaagatgctg aagatcagtt gggtgcagca aactattaac tggcgaacta
     6241 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6301 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6361 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6421 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6481 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6541 ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     6781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6841 gttctttaat agtggactct tgttccaaac ttgaacaaca ctcaacccta tctcgggcta
     6901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6961 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct
     7021 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7081 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7141 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7201 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7261 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7321 ctacatacct cgctctgcta atcctgttac cagtggggca tttgagaagc acacggtcac
     7381 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
     7441 accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc
     7501 ttattatcac ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa
     7561 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     7621 cgacatggaa gccatcacag acggcatgat gaacctgaat cgccagcggc atcagcacct
     7681 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     7741 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     7801 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     7861 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     7921 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     7981 gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa
     8041 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     8101 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     8161 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     8221 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     8281 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     8341 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     8401 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
     8461 acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc
     8521 agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga
     8581 catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg
     8641 aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata
     8701 caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     8761 agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa
     8821 cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag
     8881 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     8941 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     9001 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     9061 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     9121 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     9181 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     9241 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     9301 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     9361 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga
     9421 taaaatattt gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     9481 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     9541 cgtagaggat ctgctcatgt ttgacagctt atc
//
