LOCUS       sequence-227424-        8598 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 8598)
  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 8598)
  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: pMevT
FEATURES             Location/Qualifiers
     source          1..8598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    116..137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(152..157,158..175,176..182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    190..206
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     195..217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     214..230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     214..230
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     249..269
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        251..269
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     298..314
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     299..315
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1079..1090)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3976..3987
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4632..4649)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4802..4888
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4980..5007
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5026..5117
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5575..6030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5662..5681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5872..5893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6586..7245)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7160..7179
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7246..7348)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7740..7757)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7874..8419)
                     /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(7954..7973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8495..8514
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
       61 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      121 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      181 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      241 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggtaccggg ccccccctcg
      301 aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggat cctctagagt
      361 cgactaggag gaatataaaa tgaaaaattg tgtcatcgtc agtgcggtac gtactgctat
      421 cggtagtttt aacggttcac tcgcttccac cagcgccatc gacctggggg cgacagtaat
      481 taaagccgcc attgaacgtg caaaaatcga ttcacaacac gttgatgaag tgattatggg
      541 taacgtgtta caagccgggc tggggcaaaa tccggcgcgt caggcactgt taaaaagcgg
      601 gctggcagaa acggtgtgcg gattcacggt caataaagta tgtggttcgg gtcttaaaag
      661 tgtggcgctt gccgcccagg ccattcaggc aggtcaggcg cagagcattg tggcgggggg
      721 tatggaaaat atgagtttag ccccctactt actcgatgca aaagcacgct ctggttatcg
      781 tcttggagac ggacaggttt atgacgtaat cctgcgcgat ggcctgatgt gcgccaccca
      841 tggttatcat atggggatta ccgccgaaaa cgtggctaaa gagtacggaa ttacccgtga
      901 aatgcaggat gaactggcgc tacattcaca gcgtaaagcg gcagccgcaa ttgagtccgg
      961 tgcttttaca gccgaaatcg tcccggtaaa tgttgtcact cgaaagaaaa ccttcgtctt
     1021 cagtcaagac gaattcccga aagcgaattc aacggctgaa gcgttaggtg cattgcgccc
     1081 ggccttcgat aaagcaggaa cagtcaccgc tgggaacgcg tctggtatta acgacggtgc
     1141 tgccgctctg gtgattatgg aagaatctgc ggcgctggca gcaggcctta cccccctggc
     1201 tcgcattaaa agttatgcca gcggtggcgt gccccccgca ttgatgggta tggggccagt
     1261 acctgccacg caaaaagcgt tacaactggc ggggctgcaa ctggcggata ttgatctcat
     1321 tgaggctaat gaagcatttg ctgcacagtt ccttgccgtt gggaaaaacc tgggctttga
     1381 ttctgagaaa gtgaatgtca acggcggggc catcgcgctc gggcatccta tcggtgccag
     1441 tggtgctcgt attctggtca cactattaca tgccatgcag gcacgcgata aaacgctggg
     1501 gctggcaaca ctgtgcattg gcggcggtca gggaattgcg atggtgattg aacggttgaa
     1561 ttaaggagga cagctaaatg aaactctcaa ctaaactttg ttggtgtggt attaaaggaa
     1621 gacttaggcc gcaaaagcaa caacaattac acaatacaaa cttgcaaatg actgaactaa
     1681 aaaaacaaaa gaccgctgaa caaaaaacca gacctcaaaa tgtcggtatt aaaggtatcc
     1741 aaatttacat cccaactcaa tgtgtcaacc aatctgagct agagaaattt gatggcgttt
     1801 ctcaaggtaa atacacaatt ggtctgggcc aaaccaacat gtcttttgtc aatgacagag
     1861 aagatatcta ctcgatgtcc ctaactgttt tgtctaagtt gatcaagagt tacaacatcg
     1921 acaccaacaa aattggtaga ttagaagtcg gtactgaaac tctgattgac aagtccaagt
     1981 ctgtcaagtc tgtcttgatg caattgtttg gtgaaaacac tgacgtcgaa ggtattgaca
     2041 cgcttaatgc ctgttacggt ggtaccaacg cgttgttcaa ctctttgaac tggattgaat
     2101 ctaacgcatg ggatggtaga gacgccattg tagtttgcgg tgatattgcc atctacgata
     2161 agggtgccgc aagaccaacc ggtggtgccg gtactgttgc tatgtggatc ggtcctgatg
     2221 ctccaattgt atttgactct gtaagagctt cttacatgga acacgcctac gatttttaca
     2281 agccagattt caccagcgaa tatccttacg tcgatggtca tttttcatta acttgttacg
     2341 tcaaggctct tgatcaagtt tacaagagtt attccaagaa ggctatttct aaagggttgg
     2401 ttagcgatcc cgctggttcg gatgctttga acgttttgaa atatttcgac tacaacgttt
     2461 tccatgttcc aacctgtaaa ttggtcacaa aatcatacgg tagattacta tataacgatt
     2521 tcagagccaa tcctcaattg ttcccagaag ttgacgccga attagctact cgcgattatg
     2581 acgaatcttt aaccgataag aacattgaaa aaacttttgt taatgttgct aagccattcc
     2641 acaaagagag agttgcccaa tctttgattg ttccaacaaa cacaggtaac atgtacaccg
     2701 catctgttta tgccgccttt gcatctctat taaactatgt tggatctgac gacttacaag
     2761 gcaagcgtgt tggtttattt tcttacggtt ccggtttagc tgcatctcta tattcttgca
     2821 aaattgttgg tgacgtccaa catattatca aggaattaga tattactaac aaattagcca
     2881 agagaatcac cgaaactcca aaggattacg aagctgccat cgaattgaga gaaaatgccc
     2941 atttgaagaa gaacttcaaa cctcaaggtt ccattgagca tttgcaaagt ggtgtttact
     3001 acttgaccaa catcgatgac aaatttagaa gatcttacga tgttaaaaaa taaggaggat
     3061 tacactatgg ttttaaccaa taaaacagtc atttctggat cgaaagtcaa aagtttatca
     3121 tctgcgcaat cgagctcatc aggaccttca tcatctagtg aggaagatga ttcccgcgat
     3181 attgaaagct tggataagaa aatacgtcct ttagaagaat tagaagcatt attaagtagt
     3241 ggaaatacaa aacaattgaa gaacaaagag gtcgctgcct tggttattca cggtaagtta
     3301 cctttgtacg ctttggagaa aaaattaggt gatactacga gagcggttgc ggtacgtagg
     3361 aaggctcttt caattttggc agaagctcct gtattagcat ctgatcgttt accatataaa
     3421 aattatgact acgaccgcgt atttggcgct tgttgtgaaa atgttatagg ttacatgcct
     3481 ttgcccgttg gtgttatagg ccccttggtt atcgatggta catcttatca tataccaatg
     3541 gcaactacag agggttgttt ggtagcttct gccatgcgtg gctgtaaggc aatcaatgct
     3601 ggcggtggtg caacaactgt tttaactaag gatggtatga caagaggccc agtagtccgt
     3661 ttcccaactt tgaaaagatc tggtgcctgt aagatatggt tagactcaga agagggacaa
     3721 aacgcaatta aaaaagcttt taactctaca tcaagatttg cacgtctgca acatattcaa
     3781 acttgtctag caggagattt actcttcatg agatttagaa caactactgg tgacgcaatg
     3841 ggtatgaata tgatttctaa aggtgtcgaa tactcattaa agcaaatggt agaagagtat
     3901 ggctgggaag atatggaggt tgtctccgtt tctggtaact actgtaccga caaaaaacca
     3961 gctgccatca actggatcga aggtcgtggt aagagtgtcg tcgcagaagc tactattcct
     4021 ggtgatgttg tcagaaaagt gttaaaaagt gatgtttccg cattggttga gttgaacatt
     4081 gctaagaatt tggttggatc tgcaatggct gggtctgttg gtggatttaa cgcacatgca
     4141 gctaatttag tgacagctgt tttcttggca ttaggacaag atcctgcaca aaatgttgaa
     4201 agttccaact gtataacatt gatgaaagaa gtggacggtg atttgagaat ttccgtatcc
     4261 atgccatcca tcgaagtagg taccatcggt ggtggtactg ttctagaacc acaaggtgcc
     4321 atgttggact tattaggtgt aagaggcccg catgctaccg ctcctggtac caacgcacgt
     4381 caattagcaa gaatagttgc ctgtgccgtc ttggcaggtg aattatcctt atgtgctgcc
     4441 ctagcagccg gccatttggt tcaaagtcat atgacccaca acaggaaacc tgctgaacca
     4501 acaaaaccta acaatttgga cgccactgat ataaatcgtt tgaaagatgg gtccgtcacc
     4561 tgcattaaat cctaagtcga cctgcaggca tgcaagcttg gctgttttgg cggatgagag
     4621 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     4681 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     4741 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     4801 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     4861 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     4921 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     4981 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     5041 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5101 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5161 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5221 aagatcagtt gggtgcagca aactattaac tggcgaacta cttactctag cttcccggca
     5281 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5341 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5401 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5461 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5521 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     5581 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5641 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5701 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5761 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5821 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5881 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5941 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6001 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     6061 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6121 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6181 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6241 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     6301 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6361 atcctgttac cagtcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     6421 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     6481 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
     6541 gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
     6601 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     6661 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     6721 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     6781 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     6841 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     6901 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     6961 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     7021 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     7081 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     7141 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     7201 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     7261 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     7321 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     7381 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     7441 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat
     7501 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag
     7561 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
     7621 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     7681 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     7741 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     7801 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     7861 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     7921 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     7981 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     8041 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     8101 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     8161 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     8221 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     8281 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     8341 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     8401 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctaggctca
     8461 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     8521 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct
     8581 catgtttgac agcttatc
//
