LOCUS       sequence-332746-        4209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUC19 with Giles integrase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4209)
  AUTHORS   Kolbe K, Bell AC, Prosser GA, Assmann M, Yang HJ, Forbes HE, 
            Gallucci S, Mayer-Barber KD, Boshoff HI, Barry Iii CE
  TITLE     Development and Optimization of Chromosomally-Integrated Fluorescent
            Mycobacterium tuberculosis Reporter Constructs.
  JOURNAL   Front Microbiol. 2020 Dec 9;11:591866. doi: 
            10.3389/fmicb.2020.591866. eCollection 2020.
  PUBMED    33362741
REFERENCE   2  (bases 1 to 4209)
  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: "Front 
            Microbiol."; date: "2020-12-9"; volume: "11:591866. doi"; pages: " 
            10.3389/fmicb.2020.591866. eCollection 2020"
COMMENT     Sequence Label: pUC19 Giles integrase
FEATURES             Location/Qualifiers
     source          1..4209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(71..93)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     193..212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     406..428
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     420..437
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     421..437
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2030..2046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2030..2046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2043..2065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2054..2070
                     /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)."
     promoter        complement(join(2078..2084,2085..2102,2103..2108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2123..2144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2261..2278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2432..3020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2512..2531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3191..3982,3983..4051))
                     /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     3814..3833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4052..4156)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
       61 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      121 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
      181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      241 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
      301 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      361 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      421 gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga cacggcctgt
      481 ccgcaccgcg gatcaggcgt tgacgatgac gggctggtcg gccacgtcgg ggagtcaggc
      541 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      601 gtaactgtca gaccaagttt actcatatat actttagatt gatttcgttg ttaacggaat
      661 gatgatcgcc gcggtgacac aatcggcgta cgcggacgcc cggacgtgac cggaggcgac
      721 cgtacaccta tcccgaccgg gcactattcg gcgtggatcc ccacgcaccc ggcgaccccc
      781 gagaggacac cgcaatgggg cgacgcggca accgcgaagg cacataccgg caccggccgg
      841 acgggcgctg ggaggcccgg ttgctctatg acgacccggt caccggtgaa cgccgccggg
      901 catcgttcta cggcaagacg aagacggcgg tacgggccaa aatgcgcgaa gctatcgacc
      961 gcctcgacgc cgacgccccg gtgcgtgacg ccgcggtgac cgtggccgat tggatgcgcc
     1021 ggtggtgcac caccaccctg ttggtctccg gccgcgccga cagcaccaaa tcgttgtacg
     1081 gcacactgtc gcgccaatac gtcgaacccg ccccgttcgg cgcgatccgg ttggaccgcc
     1141 tgcgcacatc cgatgtcgag gcgttggtgc tgcgccttca acaggccacc cgcccgggcc
     1201 gcgacgggac gccggtgcgc gcggtcgccg acagcacgat ccggtcgacc tacaccgtgt
     1261 tgcgccaggc gctcgacggc gcggtgcgcg acggcctgat ggcgcgcaac cctgcggcgg
     1321 ccatgaagcg gcccgggttg tcgcgcccgg aagtccggcg gatcaacgac gcgcagttgg
     1381 ccgacctgtt ggccgccacc gccggatccc gctaccaccg cgccctgttg ctgatcgcgg
     1441 cgaccggact gcgcgccggg gaggcgttgg cccttcggtg ggacgccgtc gacctcgacg
     1501 ccgggtcgct gcgcgtggtg gccaccgtcg gccgcgtcga gcgtcgcctg cgcctcacga
     1561 cgccgaagac ggaacgatcc cgccggacgg tgccgttgac gccggacatg gtgtcgatgc
     1621 tgcgtcggca ccgcgccgac cagttagctg aacgcctgcg cgcggccaac gtgtgggagg
     1681 accacgggtt ggtgttcccg tcggcgttcg gccgcccggt ggatccgcgc aacctgttgc
     1741 gggtggtgga gcaggccgcc gcgcaggttg gcctcgaggg cgtcggcgtg cacaccctgc
     1801 gtcacgccgc ggcggtggcg atgctcgagg cgggcatcca catccgcgcg gtcgccgaca
     1861 tcctcgggca ttcgtcggtg gcggtcacgg gggacatcta cggccacacg tccgacgcga
     1921 ccgcgcgggc cgccgtcgac gtgttgtcgg cgcgactgcg gatcgcggac ggcagcggac
     1981 agtcttaggg tacccgatac cgtcggcttc caagcttggc gtaatcatgg tcatagctgt
     2041 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2101 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2161 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2221 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3181 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3781 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4141 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4201 ttatcatga
//
