LOCUS       sequence-237789-        2728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TA-cloning vector of PCR-products.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2728)
  AUTHORS   Motohashi K
  TITLE     A novel series of high-efficiency vectors for TA cloning and 
            blunt-end cloning of PCR products.
  JOURNAL   Sci Rep. 2019 Apr 23;9(1):6417. doi: 10.1038/s41598-019-42868-6.
  PUBMED    31015513
REFERENCE   2  (bases 1 to 2728)
  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: "Sci Rep."; 
            date: "2019-04-23"; volume: "9(1):6417. doi"; pages: " 
            10.1038/s41598-019-42868-6"
COMMENT     Sequence Label: pCRT
FEATURES             Location/Qualifiers
     source          1..2728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..20)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        88..192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(193..261,262..1053)
                     /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     complement(411..430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1224..1812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1713..1732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1966..1983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2100..2121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2136..2141,2142..2159,2160..2166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2174..2190
                     /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     2179..2201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2198..2214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2198..2214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2210..2575
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPQYLYGDASVSIQVLGDPLESTCRHASLALAVVLQ
                     RRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCG
                     ISHRIWCTLSTICSDAA"
     primer_bind     complement(2326..2343)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2326..2342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2335..2357)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2551..2570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2670..2692
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
       61 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      121 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      181 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      241 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      301 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      361 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      421 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      481 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      541 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      601 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      661 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      721 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
      781 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
      841 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      901 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      961 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1021 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1081 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1141 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1921 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1981 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2041 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2101 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2161 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     2221 tacgaattcg agctcggtac cccaatactt gtatggagac gctagcgtct ccatacaagt
     2281 attgggggat cctctagagt cgacctgcag gcatgcaagc ttggcactgg ccgtcgtttt
     2341 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2401 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2461 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2521 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2581 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2641 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2701 gtcatcaccg aaacgcgcga gacgaaag
//
