LOCUS       sequence-153404-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli BW25113 + JBEI-12056 (pBbA5c-MevTo-BBa1002-pTrc-MKco-PMKco, 
            JBx_041105) + JBEI-9348 (pTrc99a-PMDsc-NudB, JBx_039365).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  AUTHORS   Kang A, George KW, Wang G, Baidoo E, Keasling JD, Lee TS
  TITLE     Isopentenyl diphosphate (IPP)-bypass mevalonate pathways for 
            isopentenol production.
  JOURNAL   Metab Eng. 2016 Mar;34:25-35. doi: 10.1016/j.ymben.2015.12.002. Epub
            2015 Dec 17.
  PUBMED    26708516
REFERENCE   2  (bases 1 to 5860)
  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: "2016-03"; volume: "34:25-35. doi"; pages: " 
            10.1016/j.ymben.2015.12.002. Epub 2015 Dec 17"
COMMENT     Sequence Label: ARK1a
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(232..237,238..254,255..261)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    269..285
                     /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     274..296
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             613..624
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2082..2099)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2252..2338
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2430..2457
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2477..2568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2569..2637,2638..3429)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2787..2806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3600..4188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4089..4108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4342..4359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4374..4514
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4440..4459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4600..4622
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4700..4777
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4778..5860
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4797..4816)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcgcaacgca attaatgtga gttagcgcga attgatctgg tttgacagct tatcatcgac
       61 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
      121 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
      181 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt
      241 aatcatccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      301 agaccatgga attcaaaaga tcttttaaga aggagatata catatgaccg tttacacagc
      361 atccgttacc gcacccgtca acatcgcaac ccttaagtat tgggggaaaa gggacacgaa
      421 gttgaatctg cccaccaatt cgtccatatc agtgacttta tcgcaagatg acctcagaac
      481 gttgacctct gcggctactg cacctgagtt tgaacgcgac actttgtggt taaatggaga
      541 accacacagc atcgacaatg aaagaactca aaattgtctg cgcgacctac gccaattaag
      601 aaaggaaatg gaatcgaagg acgcctcatt gcccacatta tctcaatgga aactccacat
      661 tgtctccgaa aataactttc ctacagcagc tggtttagct tcctccgctg ctggctttgc
      721 tgcattggtc tctgcaattg ctaagttata ccaattacca cagtcaactt cagaaatatc
      781 tagaatagca agaaaggggt ctggttcagc ttgtagatcg ttgtttggcg gatacgtggc
      841 ctgggaaatg ggaaaagctg aagatggtca tgattccatg gcagtacaaa tcgcagacag
      901 ctctgactgg cctcagatga aagcttgtgt cctagttgtc agcgatatta aaaaggatgt
      961 gagttccact cagggtatgc aattgaccgt ggcaacctcc gaactattta aagaaagaat
     1021 tgaacatgtc gtaccaaaga gatttgaagt catgcgtaaa gccattgttg aaaaagattt
     1081 cgccaccttt gcaaaggaaa caatgatgga ttccaactct ttccatgcca catgtttgga
     1141 ctctttccct ccaatattct acatgaatga cacttccaag cgtatcatca gttggtgcca
     1201 caccattaat cagttttacg gagaaacaat cgttgcatac acgtttgatg caggtccaaa
     1261 tgctgtgttg tactacttag ctgaaaatga gtcgaaactc tttgcattta tctataaatt
     1321 gtttggctct gttcctggat gggacaagaa atttactact gagcagcttg aggctttcaa
     1381 ccatcaattt gaatcatcta actttactgc acgtgaattg gatcttgagt tgcaaaagga
     1441 tgttgccaga gtgattttaa ctcaagtcgg ttcaggccca caagaaacaa acgaatcttt
     1501 gattgacgca aagactggtc taccaaagga ataaggatct tttaagaagg agatatacat
     1561 atggaggata aagtgtataa gcgtcccgtt tcgatcttag tggtcatcta cgcacaagat
     1621 acgaaacggg tgctgatgtt gcagcggcgt gacgatcccg atttctggca gtcggtaacc
     1681 ggcagcgtgg aagagggtga aaccgcgccg caagctgcca tgcgcgaagt aaaggaagag
     1741 gtcaccattg atgttgtcgc tgaacaactg accttaattg actgtcagcg cacggtagag
     1801 tttgaaattt tttcacattt acgtcatcgc tatgcgccgg gcgtgacgcg taatacggaa
     1861 tcatggttct gtcttgcgct tccgcacgag cggcagatcg ttttcactga acatctggct
     1921 tacaagtggc ttgatgcgcc tgctgcggcg gcgctcacta agtcctggag caaccggcag
     1981 gcgattgaac agtttgtaat taacgctgcc tgaggatcca aactcgagta actgcaggca
     2041 tgcaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     2101 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     2161 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     2221 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2281 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2341 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     2401 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     2461 gtttctacaa actctttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tacagcaatg
     3121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3721 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4261 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4321 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4381 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     4441 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     4501 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4561 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4621 ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga
     4681 agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg
     4741 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     4801 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     4861 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     4921 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     4981 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     5041 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     5101 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     5161 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     5221 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     5281 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     5341 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     5401 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     5461 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     5521 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     5581 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgtcaaccac
     5641 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     5701 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     5761 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5821 gctggcacga caggtttccc gactggaaag cgggcagtga
//
