LOCUS       sequence-74299-e        3957 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 3957)
  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 3957)
  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: pACM4
FEATURES             Location/Qualifiers
     source          1..3957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     520..539
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(559..636)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     842..861
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    971..995
                     /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             1010..1032
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1106..1150
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1189..1207)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1203..1250
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1376..2035)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1950..1969
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2036..2138)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2530..2547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2664..3208)
                     /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(2744..2763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3433..3957,1..558))
                     /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 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
       61 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
      121 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
      181 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
      241 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
      301 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
      361 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
      421 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
      481 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
      541 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
      601 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
      661 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
      721 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
      781 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
      841 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
      901 cgatgcgtcc ggcgtagcct aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac
      961 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     1021 ttaagaagga gatatacata tggcagatct caattggata tcggccggcc acgcgatcgc
     1081 tgacgtcggt accctcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
     1141 catggactcg tctactagtc gcagcttaat taacctaaac tgctgccacc gctgagcaat
     1201 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctagcgaaag
     1261 gaggagtcga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
     1321 ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     1381 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     1441 agccatcaca gacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     1501 gcgtataata tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     1561 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     1621 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     1681 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     1741 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     1801 ctttcattgc catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa
     1861 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     1921 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     1981 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2041 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     2101 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
     2161 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
     2221 cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga
     2281 tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc
     2341 tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc
     2401 aagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
     2461 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
     2521 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     2581 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     2641 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     2701 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2761 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     2821 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     2881 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     2941 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     3001 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     3061 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     3121 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     3181 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt
     3241 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
     3301 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     3361 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     3421 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     3541 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     3601 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     3661 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
     3721 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     3781 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     3841 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     3901 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcc
//
