LOCUS       sequence-74298-e        3810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ePathBrick vector for pathway engineering in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3810)
  AUTHORS   Xu P, Vansiri A, Bhan N, Koffas MA
  TITLE     ePathBrick: a synthetic biology platform for engineering metabolic 
            pathways in E. coli.
  JOURNAL   ACS Synth Biol. 2012 Jul 20;1(7):256-66. doi: 10.1021/sb300016b. 
            Epub 2012 May 4.
  PUBMED    23651248
REFERENCE   2  (bases 1 to 3810)
  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"; doi: "10.1021/sb300016b";
            journalName: "ACS Synth Biol"; date: "2012-07-20- 20"; volume: "1"; 
            issue: "7"; pages: "256-66"
COMMENT     Sequence Label: pCDM4
FEATURES             Location/Qualifiers
     source          1..3810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     519..538
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(558..635)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     841..860
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     951..970
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        951..969
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    970..994
                     /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)."
     RBS             1009..1031
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1105..1149
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1188..1206)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1202..1249
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1406..2197)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2198..2289)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2337..3075)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(join(3285..3810,1..557))
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
       61 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
      121 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
      181 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
      241 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
      301 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
      361 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
      421 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
      481 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
      541 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
      601 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
      661 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
      721 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
      781 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
      841 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
      901 gatgcgtccg gcgtagccta ggatcgagat cgatctcgat cccgcgaaat taatacgact
      961 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     1021 taagaaggag atatacatat ggcagatctc aattggatat cggccggcca cgcgatcgct
     1081 gacgtcggta ccctcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac
     1141 atggactcgt ctactagtcg cagcttaatt aacctaaact gctgccaccg ctgagcaata
     1201 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tagcgaaagg
     1261 aggagtcgac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     1321 ctatttaacg accctgccct gaaccgacga ccgggtcatc gtggccggat cttgcggccc
     1381 ctcggcttga acgaattgtt agacattatt tgccgactac cttggtgatc tcgcctttca
     1441 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
     1501 caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     1561 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     1621 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     1681 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     1741 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     1801 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     1861 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     1921 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     1981 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     2041 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     2101 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     2161 gttgagtcga tacttcggcg atcaccgctt ccctcatact cttccttttt caatattatt
     2221 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2281 ataaacaaat agccagctca ctcggtcgct acgctccggg cgtgagactg cggcgggcgc
     2341 tgcggacaca tacaaagtta cccacagatt ccgtggataa gcaggggact aacatgtgag
     2401 gcaaaacagc agggccgcgc cggtggcgtt tttccatagg ctccgccctc ctgccagagt
     2461 tcacataaac agacgctttt ccggtgcatc tgtgggagcc gtgaggctca accatgaatc
     2521 tgacagtacg ggcgaaaccc gacaggactt aaagatcccc accgtttccg gcgggtcgct
     2581 ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc cgcctttctc
     2641 ccttacggga agtgtggcgc tttctcatag ctcacacact ggtatctcgg ctcggtgtag
     2701 gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc
     2761 ttatccggta actgttcact tgagtccaac ccggaaaagc acggtaaaac gccactggca
     2821 gcagccattg gtaactggga gttcgcagag gatttgttta gctaaacacg cggttgctct
     2881 tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga tttggttgct gtgctctgcg
     2941 aaagccagtt accacggtta agcagttccc caactgactt aaccttcgat caaaccacct
     3001 ccccaggtgg ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa aggatctcaa
     3061 gaagatcctt tgatcttttc tactgaaccg ctctagattt cagtgcaatt tatctcttca
     3121 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
     3181 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     3241 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     3301 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3361 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     3421 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     3481 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     3541 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     3601 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     3661 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     3721 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     3781 agacgcgccg agacagaact taatgggccc
//
