LOCUS       sequence-174435-        2749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for ObLiGaRe/ZFN mediated targeting to CDX2 exon1 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2749)
  AUTHORS   Pinto R, Hansen L, Hintze J, Almeida R, Larsen S, Coskun M, Davidsen
            J, Mitchelmore C, David L, Troelsen JT, Bennett EP
  TITLE     Precise integration of inducible transcriptional elements (PrIITE) 
            enables absolute control of gene expression.
  JOURNAL   Nucleic Acids Res. 2017 May 4. doi: 10.1093/nar/gkx371.
  PUBMED    28472465
REFERENCE   2  (bases 1 to 2749)
  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: "Nucleic 
            Acids Res. 2017 May 4. doi: 10.1093/nar/gkx371."
COMMENT     Sequence Label: pCDX2 ObLiGaRe Donor vector/EPB64
FEATURES             Location/Qualifiers
     source          1..2749
                     /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"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(435..453)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(436..453)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(573..589)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(573..589)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(586..608)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    597..613
                     /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(621..627,628..645,646..651))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    666..687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(804..821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(975..1563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1055..1074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1734..2549)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1734..2549)
                     /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
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2457..2476
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2550..2654)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2722..2740
                     /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 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtaaccg ccgtagtccg ggtaccgcag
      421 ggggctgacg aagttctata gtgtcaccta aatcttaatt aactcgagga tatcgtttaa
      481 acaagcttgt cgacgctagc aggccggcgg ataactagct gatcgcggaa ccgccgtagt
      541 ccgggtaccg cagggggctg acgaagttca tggtcatagc tgtttcctgt gtgaaattgt
      601 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      661 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      721 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      781 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      841 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      901 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1381 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1681 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttagaaa
     1741 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1801 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1861 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1921 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1981 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2041 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2101 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2161 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2221 aatacctgga atgctgtttt cccagggatc gcagtggtga gtaaccatgc atcatcagga
     2281 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2341 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2401 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2461 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     2521 caagacgttt cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt
     2581 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2641 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     2701 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
