LOCUS       sequence-7056-ex        6028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conditional gene targeting vector; contain cloning sites for 5′ and 
            3′ homology arms, and a central targeting region flanked by LoxP 
            sites, and a neomycin cassette flanked by FRT sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6028)
  TITLE     pEZ-Frt-lox-DT
REFERENCE   2  (bases 1 to 6028)
  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"
COMMENT     Sequence Label: pEZ-Frt-lox-DT
FEATURES             Location/Qualifiers
     source          1..6028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    33..66
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(95..142)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     222..238
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(603..622)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1214..1233
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1351..1399
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    complement(1431..1478)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    1513..1546
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1602..1826
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1815..1832
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     intron          1900..1965
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(2042..2614)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     promoter        complement(2641..3140)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2689..2708)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     3082..3105
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(3170..3189)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3171..3189)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3199..3215)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3357..3812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3444..3463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3654..3675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3838..3942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3943..4011,4012..4803)
                     /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(4161..4180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4974..5562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5463..5482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5716..5733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5850..5871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5886..5891,5892..5909,5910..5916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5924..5940
                     /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     5929..5951
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5948..5964
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5948..5964
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5983..6003
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5985..6003
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tagatatcga tggccatagc ggccgcggat ccataacttc gtataatgta tgctatacga
       61 agttatcctt tcgagggagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
      121 ggaacttcgg aataggaact tcaagatccc cctggcgaaa gggggatgtg ctgcaaggcg
      181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
      241 attcgagctc tcccatatgg tcgagcagtg tggttttcaa gaggaagcaa aaagcctctc
      301 cacccaggcc tggaatgttt ccacccaatg tcgagcagtg tggttttgca agaggaagca
      361 aaaagcctct ccacccaggc ctggaatgtt tccacccaat gtcgagcaaa ccccgcccag
      421 cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc
      481 cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct gcagccaata
      541 tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      601 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      661 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      721 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
      781 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
      841 cggggcagga tctcctgtgc atctcacctt gctcctgccg agaaagtatc catcatggct
      901 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
      961 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     1021 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     1081 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     1141 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     1201 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     1261 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     1321 cgccttcttg acgagttctt ctgaggggat cggcaataaa aagacagaat aaaacgcacg
     1381 ggtgttgggt cgtttgttcg gatccgtcga gggagcttca aaagcgctct gaagttccta
     1441 tactttctag agaataggaa cttcggaata ggaacttcaa gatccccctg gcgaaagggg
     1501 gatgtggtcg acataacttc gtataatgta tgctatacga agttataagc ttgcggccgg
     1561 ccgccacctc gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
     1621 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     1681 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     1741 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     1801 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctag gatctgcatt
     1861 ccaccactgc tcccattcat cagttccata ggttggaatc taaaatacac aaacaattag
     1921 aatcagtagt ttaacacatt atacacttaa aaattttata tttaccttag agctttaaat
     1981 ctctgtaggt agtttgtcca attatgtcac accacagaag taaggttcct tcacaaagag
     2041 atcgcctgac acgatttcct gcacaggctt gagccatata ctcatacatc gcatcttggc
     2101 cacgttttcc acgggtttca aaattaatct caagttctac gcttaacgct ttcgcctgtt
     2161 cccagttatt aatatattca acgctagaac tcccctcagc gaagggaagg ctgagcacta
     2221 cacgcgaagc accatcaccg aaccttttga taaactcttc cgttccgact tgctccatca
     2281 acggttcagt gagacttaaa cctaactctt tcttaatagt ttcggcatta tccactttta
     2341 gtgcgagaac cttcgtcagt cctggatacg tcactttgac cacgcctcca gcttttccag
     2401 agagcgggtt ttcattatct acagagtatc ccgcagcgtc gtatttattg tcggtactat
     2461 aaaacccttt ccaatcatcg tcataatttc cttgtgtacc agattttggc ttttgtatac
     2521 ctttttgaat ggaatctaca taaccaggtt tagtcccgtg gtacgaagaa aagttttcca
     2581 tcacaaaaga tttagaagaa tcaacaacat catcaggatc catggcgagg acctgcaggt
     2641 cgaaaggccc ggagatgagg aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt
     2701 gcagaatgcc gggcctccgg aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct
     2761 gagcccgccc ccggacccac cccttcccag cctctgagcc cagaaagcga aggagcaaag
     2821 ctgctattgg ccgctgcccc aaaggcctac ccgcttccat tgctcagcgg tgctgtccat
     2881 ctgcacgaga ctagtgagac gtgctacttc catttgtcac gtcctgcacg acgcgagctg
     2941 cggggcgggg gggaacttcc tgactagggg aggagtagaa ggtggcgcga aggggccacc
     3001 aaagaacgga gccggttggc gcctaccggt ggatgtggaa tgtgtgcgag gccagaggcc
     3061 acttgtgtag cgccaagtgc ccagcggggc tgctaaagcg catgctccag actgccttgg
     3121 gaaaagcgcc tcccctaccc ggtagaattc gatgtggagc tccaattcgc cctatagtga
     3181 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3241 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3301 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     3361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3421 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3541 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3781 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     3841 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3901 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3961 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4021 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4081 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4141 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4201 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4261 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4321 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4381 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4441 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4501 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4561 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4621 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4681 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4741 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4801 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4861 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4921 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5101 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     5161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5701 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     5761 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5821 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     5881 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     5941 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     6001 agggaacaaa agctggtcga gatctagc
//
