LOCUS       sequence-447549-        3024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RBS_L2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 144 to 178)
  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 3024)
  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-TL9
FEATURES             Location/Qualifiers
     source          1..3024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..57
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(61..104)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    107..126
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        144..178
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     CDS             239..946
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     primer_bind     complement(263..291)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     886..906
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      962..1033
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1049..1076
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1104..1161
                     /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(1185..1202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1356..1944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1436..1455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2127..2221)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2242..2901)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2816..2835
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga
       61 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
      121 ttctagaacg tctcaatctc tatttgacgg ctagctcagt cctaggtaca gtgctagcgg
      181 tctcagtaca gatctaataa ttttgtttaa ctttaaaagg atacatatga gaccgcgtaa
      241 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
      301 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
      361 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
      421 tttcggttat ggtgttcaat gctttgcgag atacccagat catatgaaac agcatgactt
      481 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
      541 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
      601 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
      661 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
      721 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
      781 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
      841 acaatctgcc ctttcgaaag atcccaacga aaagagagat cacatggtcc ttcttgagtt
      901 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataat cgaccaggca
      961 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1021 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     1081 tatgtttgcc agagacgctg cagtccggca aaaaagggca aggtgtcacc accctgccct
     1141 ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct cactgactcg
     1201 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1261 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1321 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac
     1381 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1441 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1501 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1561 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1621 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1681 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1741 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1801 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1861 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1921 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1981 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2041 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2101 acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     2161 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     2221 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     2281 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     2341 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     2401 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     2461 ggctgacacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     2521 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     2581 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     2641 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     2701 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     2761 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     2821 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     2881 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     2941 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc
     3001 gatcaactcg agtgccacct gacg
//
