LOCUS       sequence-74282-e        5203 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 5203)
  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 5203)
  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: pETM6
FEATURES             Location/Qualifiers
     source          1..5203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             138..182
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(221..239)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      235..282
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      326..781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     623..644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(896..1687,1688..1756))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1519..1538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1757..1849)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1930..2518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2419..2438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2672..2689
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2704..2846
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2772..2791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2932..2954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2948..3139)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3711..4793)
                     /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"
     primer_bind     4755..4774
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4794..4871)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5077..5096
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(5187..5203,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5187..5203,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatataca tatggcagat ctcaattgga tatcggccgg ccacgcgatc gctgacgtcg
      121 gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact
      181 cgtctactag tcgcagctta attaacctaa actgctgcca ccgctgagca ataactagca
      241 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctagcgaa aggaggagtc
      301 gactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      721 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
      781 tctggcggca cgatggcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1741 aatgttgaat actcatactc ttcctttttc aatcatgatt gaagcattta tcagggttat
     1801 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggtcatgac
     1861 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1921 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1981 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2041 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2101 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2161 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2221 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2281 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2341 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2401 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2461 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2521 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2581 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2641 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2701 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2761 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2821 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2881 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2941 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3001 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3061 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3121 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     3181 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     3241 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3301 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3361 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3421 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3481 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3541 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgctag
     3601 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     3661 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     3721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3781 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3841 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3901 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     3961 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     4021 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4081 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4141 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4201 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4261 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4321 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4381 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4441 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4501 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4561 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4621 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4681 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4741 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4801 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4861 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     4921 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     4981 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     5041 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     5101 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agcctaggat
     5161 cgagatcgat ctcgatcccg cgaaattaat acgactcact ata
//
