LOCUS       sequence-141169-        3040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RBS_H1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 2450 to 2484)
  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 3040)
  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-TL1
FEATURES             Location/Qualifiers
     source          1..3040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..172
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      228..299
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      315..342
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    365..385
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      386..443
                     /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(467..484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(638..1226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(718..737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1409..1503)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1524..2183)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2098..2117
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2345..2363
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2367..2410)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2413..2432
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        2450..2484
                     /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             join(2545..3040,1..212)
                     /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(2569..2597)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
       61 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
      121 cgaaagatcc caacgaaaag agagatcaca tggtccttct tgagtttgta acagctgctg
      181 ggattacaca tggcatggat gaactataca aataatcgac caggcatcaa ataaaacgaa
      241 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
      301 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatatg tttgccagag
      361 acggtactag tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc
      421 ctttttcttt aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact
      481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      541 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc cgcccccctg
      661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1201 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1321 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1381 aaacttggtc tgacagctcg aggcttggat tctcaccaat aaaaaacgcc cggcggcaac
     1441 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     1501 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     1561 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     1621 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     1681 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     1741 ttggctgaca cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     1801 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     1861 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     1921 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     1981 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     2041 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     2101 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     2161 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     2221 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     2281 ccgatcaact cgagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     2341 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
     2401 ggatgatttc tggaattcgc ggccgcttct agaacgtctc aatctctatt tgacggctag
     2461 ctcagtccta ggtacagtgc tagcggtctc agtacagatc taataatttt gtttaacttt
     2521 ggggggatac atatgagacc gcgtaaagga gaagaacttt tcactggagt tgtcccaatt
     2581 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     2641 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
     2701 gttccatggc caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac
     2761 ccagatcata tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     2821 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
     2881 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     2941 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
     3001 gacaaacaaa agaatggaat caaagttaac ttcaaaatta
//
