LOCUS       sequence-334795-        3390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  THe donor vector for the KLF7 gene locus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3390)
  AUTHORS   Yoshino Y, Endo S, Chen Z, Qi H, Watanabe G, Chiba N
  TITLE     Evaluation of site-specific homologous recombination activity of 
            BRCA1 by direct quantitation of gene editing efficiency.
  JOURNAL   Sci Rep. 2019 Feb 7;9(1):1644. doi: 10.1038/s41598-018-38311-x.
  PUBMED    30733539
REFERENCE   2  (bases 1 to 3390)
  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: "2019-02-7"; volume: "9(1):1644. doi"; pages: " 
            10.1038/s41598-018-38311-x"
COMMENT     Sequence Label: pBS-KLF7-200-GFPfr1
FEATURES             Location/Qualifiers
     source          1..3390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(14..469)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(151..172)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     363..382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     596..618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     610..627
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     611..627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     637..656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     680..696
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     681..697
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1160..1176)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1160..1176)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1213..1233)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1213..1231)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1252..1268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1252..1268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1265..1287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1276..1292
                     /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(1300..1306,1307..1324,1325..1330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1345..1366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1483..1500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1654..2242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1734..1753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2413..3204,3205..3273))
                     /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     3036..3055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3274..3378)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
       61 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
      121 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      181 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      241 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      301 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      361 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      421 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
      481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      601 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
      661 ttgggtaccg ggccccccct cgaggtcgac ggtatcgata agctttccag tggccctttt
      721 gttttgtttt gtttcagtca actaaaaagg aaaaaaaaaa atcaatgcag gagagggaga
      781 gaggaggtga gagaggagcg agtgagtggg gtggatggag agaggcatcc agcgtgtaca
      841 gtgcagacga ctgccaggaa aaggggactt ctccacggga gtaacaattc ccttccaggg
      901 ctctagtcct gctggagttc gtgaccgcaa gggggatgca aactcactga cacgaagctg
      961 ccatctattt ctgcagcgct gttcgctcct aatgcaatct ttgctcttta ttttgggagg
     1021 ttgcattttt ttctctcgcg atcgcttctc ttcccccccc ttcccttccc tccccacccc
     1081 ccaccccatc cttgctagtt tgtagtcttc cccctccctt ttctcctcaa gcttgatatc
     1141 gaattcctgc agcccggggg atccactagt tctagagcgg ccgccaccgc ggtggagctc
     1201 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     1261 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1321 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3361 acaaataggg gttccgcgca catttccccg
//
