LOCUS       sequence-74301-e        3777 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 3777)
  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 3777)
  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: pRSM3
FEATURES             Location/Qualifiers
     source          1..3777
                     /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(1385..2200)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1385..2200)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2108..2127
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2201..2292)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2308..3057)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2318..2335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2558..2577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3253..3777,1..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 caagctgacg accgggtctc cgcaagtggc acttttcggg gaaatgtgcg
     1321 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     1381 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1441 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1501 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1561 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1621 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     1681 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1741 attgcgcctg agcgagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa
     1801 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1861 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     1921 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     1981 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2041 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2101 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2161 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat actcttcctt tttcaatatt
     2221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2281 aaaataaaca aataggcatg cagcgctctt ccgcttcctc gctcactgac tcgctacgct
     2341 cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa agcggtaata cggttatcca
     2401 cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag
     2461 ccaacgccgc aggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2521 cgctcaagcc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2581 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2641 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgttggta tctcagttcg
     2701 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2761 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2821 ctggcagcag ccattggtaa ctgatttaga ggactttgtc ttgaagttat gcacctgtta
     2881 aggctaaact gaaagaacag attttggtga gtgcggtcct ccaacccact taccttggtt
     2941 caaagagttg gtagctcagc gaaccttgag aaaaccaccg ttggtagcgg tggtttttct
     3001 ttatttatga gatgatgaat caatcggtct atcaagtcaa cgaacagcta ttccgttact
     3061 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
     3121 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     3181 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     3361 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     3421 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     3481 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
     3541 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     3601 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     3661 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     3721 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcc
//
