LOCUS       sequence-371220-        3987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SCB-GFP E. coli reporter plasmid. Encodes repressor ScbR and ScbAp 
            promoter upstream of GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 130 to 164)
  AUTHORS   Biarnes-Carrera M, Lee CK, Nihira T, Breitling R, Takano E
  TITLE     Orthogonal Regulatory Circuits for Escherichia coli Based on the 
            gamma-Butyrolactone System of Streptomyces coelicolor.
  JOURNAL   ACS Synth Biol. 2018 Apr 20;7(4):1043-1055. doi: 
            10.1021/acssynbio.7b00425. Epub 2018 Mar 19.
  PUBMED    29510026
REFERENCE   2  (bases 1 to 3987)
  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/acssynbio.7b00425"; journalName: "ACS Synth Biol"; date: 
            "2018-04-20- 20"; volume: "7"; issue: "4"; pages: "1043-1055"
COMMENT     Sequence Label: pTE1069
FEATURES             Location/Qualifiers
     source          1..3987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..73
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(77..120)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        130..164
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             171..182
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      910..981
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      997..1024
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1162..1173
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1180..1893
                     /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="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1210..1238)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1833..1853
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1947..2018
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2034..2061
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2083..2140
                     /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(2164..2181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2415..2434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3106..3200)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3221..3880)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3795..3814
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
       61 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
      121 tggctgcagt tgacggctag ctcagtccta ggtacagtgc tagcggatcc aaagaggaga
      181 aagaattcat ggccaagcag gaccgggcga tccgcacgcg gcagacgatc ctggacgccg
      241 cggcgcaggt cttcgagaag cagggctacc aagctgccac gatcacggag atcctcaagg
      301 tggccggggt gaccaaggga gccctctact tccacttcca gtccaaggaa gaactggcgc
      361 tgggcgtctt cgacgcccag gaaccaccac aggccgttcc ggagcaaccc ctccggctgc
      421 aagaactcat cgacatgggc atgttgttct gtcaccgctt gcgcacgaac gtcgtggccc
      481 gggccggcgt gcgcctctcc atggaccagc aggcgcacgg tctcgatcgc cgaggaccct
      541 tccgtcgctg gcacgagaca ctcctgaagc tgctgaacca ggccaaggag aacggtgagt
      601 tgctgcccca tgtggtcacc accgactcgg ccgatctcta cgtgggcacg ttcgccggga
      661 tacaggtcgt gtcccagacg gtcagcgact accaggacct cgaacaccgc tacgcgctgc
      721 tgcagaagca catcctgccc gccatcgcgg ttccctccgt gctggccgcg ctcgatctct
      781 ccgaggagcg cggagcacgc ctcgcggccg aactggcacc gaccgggaag gaccgtcgtc
      841 gccgtcgccg ttgataataa gcggccgcca ctgatagtgc tagtgtagat cactactaga
      901 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      961 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     1021 tgcgtttata ggtacctact agagaggttc tgttaagtaa ctgaacccaa tgtcgttagt
     1081 gacgcttacc tcttaagagg tcactgacct aacaaagctt tagaaaaaaa accgctcagt
     1141 ctgtatctta acgttactag taaagaggag aaatagcata tgcgtaaagg agaagaactt
     1201 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1261 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1321 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt cggttatggt
     1381 gttcaatgct ttgcgagata cccagatcac atgaaacagc atgacttttt caagagtgcc
     1441 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1501 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1561 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
     1621 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt
     1681 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     1741 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     1801 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct
     1861 gggattacac atggcatgga tgaactatac aaataataat ctagacactg atagtgctag
     1921 tgtagatcac tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1981 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     2041 cttcgggtgg gcctttctgc gtttataccc gggtactaga ggtccggcaa aaaagggcaa
     2101 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc
     2161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2341 caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2521 gctttctcat agctcacgct gtaagtatct cagttcggtg taggtcgttc gctccaagct
     2581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2941 ttctacgggg tcagacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3061 ctaaagtata tatgagtaaa cttggtctga cagctcgagg cttggattct caccaataaa
     3121 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     3181 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     3241 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     3301 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     3361 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     3421 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     3481 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     3541 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     3601 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     3661 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     3721 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     3781 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     3841 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     3901 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     3961 tggaacctct tacgtgcccg atcaact
//
