LOCUS       sequence-447502-        2206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 promoter and RBS  - derived from pET3a (5' CTAT/3' CATA) fusion).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2206)
  AUTHORS   Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont
            PS
  TITLE     EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2016 May 2.
  PUBMED    27096716
REFERENCE   2  (bases 1 to 2206)
  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: "ACS Synth 
            Biol. 2016 May 2."
COMMENT     Sequence Label: pBP-T7_RBS
FEATURES             Location/Qualifiers
     source          1..2206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..82
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(86..129)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    132..151
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     191..210
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        191..209
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    210..234
                     /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             249..271
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      311..368
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(392..409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(563..1151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(643..662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1334..1428)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1449..2108)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2023..2042
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgatcaactc gagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
       61 aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
      121 gatgatttct ggaattcgcg gccgcttcta gaatctagag acgcatatgg gtctcactat
      181 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
      241 gaaataattt tgtttaactt taagaaggag atataccata agagaccgca tgccgtctca
      301 ttagctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac
      361 cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg
      421 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      481 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      541 aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa
      601 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      661 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      721 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      781 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      841 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      901 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      961 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1021 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1081 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1141 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1201 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1261 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1321 gctcgaggct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     1381 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     1441 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     1501 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     1561 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     1621 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgacacgaaa
     1681 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     1741 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     1801 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     1861 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
     1921 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     1981 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     2041 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     2101 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     2161 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcc
//
