LOCUS       sequence-178357-        5852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integration-proficient plasmid vector for mycobacteria, contains the
            attP and int region from Tweety genomic DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5852)
  AUTHORS   Pham TT, Jacobs-Sera D, Pedulla ML, Hendrix RW, Hatfull GF
  TITLE     Comparative genomic analysis of mycobacteriophage Tweety: 
            evolutionary insights and construction of compatible site-specific 
            integration vectors for mycobacteria.
  JOURNAL   Microbiology. 2007 Aug;153(Pt 8):2711-23.
  PUBMED    17660435
REFERENCE   2  (bases 1 to 5852)
  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: 
            "Microbiology."; date: "2007-08"; volume: "153(Pt 8)"; pages: 
            "2711-23"
COMMENT     Sequence Label: pTTP1B
FEATURES             Location/Qualifiers
     source          1..5852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(187..642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(324..345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     536..555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     840..862
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     854..871
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     855..871
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2720..2739)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2731..3546
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKFLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2804..2823)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(3632..3648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3632..3648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3645..3667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3656..3672
                     /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(3680..3686,3687..3704,3705..3710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3725..3746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3863..3880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4034..4621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4792..5583,5584..5652))
                     /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     5415..5434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5653..5757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5825..5843
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      241 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
      301 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      361 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      421 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      481 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      541 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      601 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
      661 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      721 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      781 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      841 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatcctc
      901 tagagtcgag atttacagtc ggtcgcctac gcaaacgtcg atgggagcag ttcggacagc
      961 ccctgcatgg cctccagatg cctcgcccga tccgcatgcg catagatccg ctgcgcatcc
     1021 acactcgcat gacccaagat ctccatacgc gtttgctcat ccacacccgc tgcgcgcagc
     1081 aatgtcgagg tggtgtgccg cgagttgtgc ggcggtaacg actcggttgg accgatcacc
     1141 ccagcagcgc ggaacacgcc acgccacacg tcgtagtccg aacggggatc gatcggcttc
     1201 ccctccttgt gccacaccaa gtcatgcgga ttgtcggtgc ggagtttctg catcgccaca
     1261 tacaacggcg gcaacaacgg aacctcacgc caaccagcgt ccgtcttcgg ccgggtgaac
     1321 aacaacgacc cctcacattc ctgatactcg aaatgcgccg gcaggtccca ccgggactgc
     1381 gggcatgccc atgcccgtgt cttcccgcaa ggccagtacg ggggtttctt gggcatacgg
     1441 tcgggccggg acagcggtga cggttcgggt agaggatccc cacagccgtg gacgcgggtt
     1501 tccgattgca actgccaagc gatggtgatc catccctgag cggggttgtc gacgtagggc
     1561 cagcgcaggc cgaggagttc cccacggcgg gcgcccgtca ggaaaccggc ggcgatccgc
     1621 accgcatccg gttcgtcgca cacctggaac gcggtgtgaa tgatgtgctg cgccacgtcc
     1681 gccgggaagc cgttgcgttt cttcttccgg tactcgggct tgtcgaccaa tgcggccaca
     1741 ttcctggtcg ccacaccctc cgctaccgca tcgtccaagg ctttctggac gatgacatgg
     1801 accagctcgg cggtgcggga ggccccgatc tcggagtgca ggtcccgcac atgctgcggg
     1861 gtgagtttgt cgatgcgttt cgcgccgagg atcgggttga tgtggttgtg gatggcggcc
     1921 cggtagtcgt tgaggacgcc ggggcggact ttacgtttgg cgtggatgtt gtcgatccag
     1981 tgcagcatcc acttctccac agttgtggat gaggtggtgg cgatgcggcc ctcttcgacg
     2041 tcgcggcgga gttgtttgag tttggccatg gcggtgttgc ggtcaacgga ggacacccat
     2101 ttgtagcggc ggttgccgtt gcggtctggg ggtagttcta ctcgtcccat ccatttgccg
     2161 tcggcgcgtt ggaagaacgc tccgtctccg cgggttctgc gtttcttagt tgccatcgtt
     2221 ttccctccca gggggtcacc ctacggttca ccctacggtg ctgcgcagca ttacgcagaa
     2281 ttgcgcagta tcgggggtct acctgcgggt ttgacaacgt ttctcctggt atgcagccta
     2341 tcaaccgctg actcttaatc agcgggtcgg gggttcgaaa ccctcacggc gcacaggtca
     2401 gaggccataa gcctcagagg ggatcaccct aaaggtaacc ctataggggt tttcactggt
     2461 cctacatgtc gtcgcgccgt cgtactctct tttcataggg gagcatgagg atccaactga
     2521 gcttgctgtg ccttcgattg agcggttgaa gcatctggag cgtcggcgga cagcgatcac
     2581 ggttcccaac gatggctgaa agaaggatga aatgactgca ctcgacctgc aggcatgcaa
     2641 gcttggccgg gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     2701 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     2761 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2821 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     2881 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2941 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     3001 cctgatgatg catggttact caccactgcg atccccggga aaacagcatt ccaggtatta
     3061 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     3121 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     3181 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     3241 aatggttggc ctgttgaaca agtctggaaa gaaatgcata agtttttgcc attctcaccg
     3301 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3361 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3421 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3481 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3541 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     3601 gacggcgccg gccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3661 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3721 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3781 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3841 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg tagtatctca gttcggtgta
     4261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5761 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5821 taaaaatagg cgtatcacga ggccctttcg tc
//
