LOCUS       sequence-116876-        5012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTrc plasmid expressing GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5012)
  AUTHORS   Sekar K, Gentile AM, Bostick JW, Tyo KE
  TITLE     N-Terminal-Based Targeted, Inducible Protein Degradation in 
            Escherichia coli.
  JOURNAL   PLoS One. 2016 Feb 22;11(2):e0149746. doi: 
            10.1371/journal.pone.0149746. eCollection 2016.
  PUBMED    26900850
REFERENCE   2  (bases 1 to 5012)
  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: "PLoS One.";
            date: "2016-02-22"; volume: "11(2):e0149746. doi"; pages: " 
            10.1371/journal.pone.0149746. eCollection 2016"
COMMENT     Sequence Label: pJB005
FEATURES             Location/Qualifiers
     source          1..5012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             426..1139
                     /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"
     CDS             426..1139
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(456..484)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1079..1099
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1366..1452
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1544..1571
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1590..1681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1682..1750,1751..2542)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1900..1919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2713..3301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3202..3221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3455..3472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3487..3627
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3553..3572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3713..3735
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3813..3890
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3891..4973
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3910..3929)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatggatcc
      421 agaggatgcg taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
      481 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
      541 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa
      601 cacttgtcac tactttcggt tatggtgttc aatgctttgc gagataccca gatcatatga
      661 aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
      721 ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc
      781 ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac
      841 acaaattgga atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga
      901 atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc gttcaactag
      961 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
     1021 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
     1081 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaaa
     1141 ggcctgcagc aaacgacgaa aactacgctg catcagttta agagctcgag atctgcagct
     1201 ggtaccatat gggaattcga agctttctag aactagtaca gaatttgcct ggcggcagta
     1261 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     1321 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     1381 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     1441 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     1501 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     1561 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
     1621 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1681 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1741 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1801 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1861 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1921 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1981 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2041 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2101 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2161 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2221 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2281 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2341 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2401 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2461 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2521 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2581 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2641 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2821 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3421 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3481 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     3541 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3601 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3661 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3721 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
     3781 cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc
     3841 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     3901 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     3961 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     4021 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     4081 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     4141 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     4201 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     4261 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     4321 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     4381 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     4441 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     4501 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     4561 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     4621 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     4681 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     4741 cgccgtcaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     4801 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     4861 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4921 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4981 gcaattaatg tgagttagcg cgaattgatc tg
//
