LOCUS       sequence-225632-        4545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for E.coli K12 fliC promoter activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4545)
  AUTHORS   Parker DJ, Demetci P, Li GW
  TITLE     Rapid Accumulation of Motility-Activating Mutations in Resting 
            Liquid Culture of Escherichia coli.
  JOURNAL   J Bacteriol. 2019 Sep 6;201(19). pii: JB.00259-19. doi: 
            10.1128/JB.00259-19. Print 2019 Oct 1.
  PUBMED    31285239
REFERENCE   2  (bases 1 to 4545)
  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: "J 
            Bacteriol."; date: "2019-09-6"; volume: "201(19). pii: JB.00259-19. 
            doi"; pages: " 10.1128/JB.00259-19. Print 2019 Oct 1"
COMMENT     Sequence Label: pGL001
FEATURES             Location/Qualifiers
     source          1..4545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             246..962
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKVEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(276..304)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     899..919
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      complement(1007..1050)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1089..1175
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1713..2663)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILHYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2711..2933
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3425..3519)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3532..4323,4324..4392))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4155..4174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4393..4497)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgatacaaaa aacccgcttc aaaaagcggg ttttttatca gacagatgta agtaattatt
       61 acaggattaa tagcgggaat aaggggcaga gaaaagagta tttcggcgac taacaaaaaa
      121 tggctgtttt tgaaaaaaat tctaaaggtt gttttacgac agacgataac agggttgacg
      181 gcgattgagc cgacgggtgg aaacccaata cgtaatcaac gacttgcaat ataggataac
      241 gaatcatgag taaagtagaa gaacttttca ctggagttgt cccaattctt gttgaattag
      301 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
      361 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa
      421 cacttgtcac tactttcgcg tatggtcttc aatgctttgc gagataccca gatcatatga
      481 aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
      541 ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc
      601 ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac
      661 acaaattgga atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga
      721 atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc gttcaactag
      781 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
      841 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
      901 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat
      961 agctagccgc atgcaagcta agaagcttgg gcccgaacaa aaactcataa aacgaaaggc
     1021 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaact ggatcctgct
     1081 agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1141 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggtacgg
     1201 gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag aatcgcagat
     1261 ccggcttcag ccggtttgcc ggctgaaagc gctatttctt ccagaattgc catgattttt
     1321 tccccacggg aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg ataagcagca
     1381 tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt ctcaggtgtt
     1441 caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta ggtgttacat
     1501 gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttgaat gcaccaaaaa
     1561 ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg catatggaca
     1621 gttttccctt tgatatgtaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg
     1681 cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg
     1741 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatac
     1801 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
     1861 aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga tgtaatggtt
     1921 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
     1981 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
     2041 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
     2101 tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca
     2161 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
     2221 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
     2281 tttatgaatt tttttaactg gaaaagataa tgcaatatct cttcactaaa aactaattct
     2341 aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc
     2401 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
     2461 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
     2521 accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
     2581 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
     2641 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
     2701 caattgagat gggctagtca atgataatta ctagtccttt tcccgggaga tctgggtatc
     2761 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
     2821 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
     2881 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
     2941 tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac
     3001 agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat
     3061 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
     3121 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
     3181 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
     3241 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
     3301 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
     3361 atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg tccctagtgc
     3421 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     3481 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc tttaccaatg
     3541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3841 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4141 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4501 atttccccga aaagtgccac ctgaccagct ggcaattccg acgtc
//
