LOCUS       Exported                3518 bp DNA     circular SYN 15-JAN-2026
DEFINITION  Parental vector for TOPO(R) cloning of blunt PCR products.
ACCESSION   .
VERSION     .
KEYWORDS    pCR-Blunt II-TOPO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  AUTHORS   Thermo Fisher (Invitrogen)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3518)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3518
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             217..576
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALKGEFC
                     RYPSHWRPLEHASRGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRN
                     SEEARTDRPSQQLRSL"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(407..425)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(432..448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             586..888
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             1237..2031
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2237..2611
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      2749..3337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttaagg gcgaattctg cagatatcca
      361 tcacactggc ggccgctcga gcatgcatct agagggccca attcgcccta tagtgagtcg
      421 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      481 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      541 cgcaccgatc gcccttccca acagttgcgc agcctatacg tacggcagtt taaggtttac
      601 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
      661 acgccggggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
      721 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
      781 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
      841 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggcatg
      901 agattatcaa aaaggatctt cacctagatc cttttcacgt agaaagccag tccgcagaaa
      961 cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc
     1021 gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt
     1081 ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag
     1141 ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca
     1201 agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     1261 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     1321 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     1381 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     1441 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     1501 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     1561 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     1621 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     1681 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     1741 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     1801 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     1861 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     1921 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     1981 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg aattattaac
     2041 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2101 atcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatagca
     2221 cgtgaggagg gccaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     2281 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     2341 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     2401 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     2461 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     2521 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     2581 cgtgcacttc gtggccgagg agcaggactg acacgtgcta aaacttcatt tttaatttaa
     2641 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2761 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2821 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2881 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2941 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3001 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3061 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3121 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3181 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3241 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3301 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3361 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3421 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3481 gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
