LOCUS       sequence-254637-        7936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human CTCF-GFP in mammalian cells through transient 
            transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7936)
  AUTHORS   Wang AJ, Han Y, Jia N, Chen P, Minden MD
  TITLE     NPM1c impedes CTCF functions through cytoplasmic mislocalization in 
            acute myeloid leukemia.
  JOURNAL   Leukemia. 2020 May;34(5):1278-1290. doi: 10.1038/s41375-019-0681-8. 
            Epub 2019 Dec 12.
  PUBMED    31831844
REFERENCE   2  (bases 1 to 7936)
  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"; doi: 
            "10.1038/s41375-019-0681-8"; journalName: "Leukemia"; date: 
            "2020-05"; volume: "34"; issue: "5"; pages: "1278-1290"
COMMENT     Sequence Label: pAcGFP-Hyg-CTCF-N1
FEATURES             Location/Qualifiers
     source          1..7936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2824..3543
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    3665..3786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3756..3775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3793..4248)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3930..3951)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4142..4161
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4275..4379
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4381..4738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4381..4572
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4404..4424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4589..4724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4746..5771
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4746..5771
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5888..5969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5997..6015)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6083..6187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6188..6256,6257..7048)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6406..6425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7234..7822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7723..7742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatg gaaggtgatg cagtcgaagc cattgtggag
      661 gagtccgaaa cttttattaa aggaaaggag agaaagactt accagagacg ccgggaaggg
      721 ggccaggaag aagatgcctg ccacttaccc cagaaccaga cggatggggg tgaggtggtc
      781 caggatgtca acagcagtgt acagatggtg atgatggaac agctggaccc cacccttctt
      841 cagatgaaga ctgaagtaat ggagggcaca gtggctccag aagcagaggc tgctgtggac
      901 gatacccaga ttataacttt acaggttgta aatatggagg aacagcccat aaacatagga
      961 gaacttcagc ttgttcaagt acctgttcct gtgactgtac ctgttgctac cacttcagta
     1021 gaagaacttc agggggctta tgaaaatgaa gtgtctaaag agggccttgc ggaaagtgaa
     1081 cccatgatat gccacaccct acctttgcct gaagggtttc aggtggttaa agtgggggcc
     1141 aatggagagg tggagacact agaacaaggg gaacttccac cccaggaaga tcctagttgg
     1201 caaaaagacc cagactatca gccaccagcc aaaaaaacaa agaaaaccaa aaagagcaaa
     1261 ctgcgttata cagaggaggg caaagatgta gatgtgtctg tctacgattt tgaggaagaa
     1321 cagcaggagg gtctgctatc agaggttaat gcagagaaag tggttggtaa tatgaagcct
     1381 ccaaagccaa caaaaattaa aaagaaaggt gtaaagaaga cattccagtg tgagctttgc
     1441 agttacacgt gtccacggcg ttcaaatttg gatcgtcaca tgaaaagcca cactgatgag
     1501 agaccacaca agtgccatct ctgtggcagg gcattcagaa cagtcaccct cctgaggaat
     1561 caccttaaca cacacacagg tactcgtcct cacaagtgcc cagactgcga catggccttt
     1621 gtgaccagtg gagaattggt tcggcatcgt cgttacaaac acacccacga gaagccattc
     1681 aagtgttcca tgtgcgatta cgccagtgta gaagtcagca aattaaaacg tcacattcgc
     1741 tctcatactg gagagcgtcc gtttcagtgc agtttgtgca gttatgccag cagggacaca
     1801 tacaagctga aaaggcacat gagaacccat tcaggggaaa agccttatga atgttatatt
     1861 tgtcatgctc ggtttaccca aagtggtacc atgaagatgc acattttaca gaagcacaca
     1921 gaaaatgtgg ccaaatttca ctgtccccac tgtgacacag tcatagcccg aaaaagtgat
     1981 ttgggtgtcc acttgcgaaa gcagcattcc tatattgagc aaggcaagaa atgccgttac
     2041 tgtgatgctg tgtttcatga gcgctatgcc ctcatccagc atcagaagtc acacaagaat
     2101 gagaagcgct ttaagtgtga ccagtgtgat tacgcttgta gacaggagag gcacatgatc
     2161 atgcacaagc gcacccacac cggggagaag ccttacgcct gcagccactg cgataagacc
     2221 ttccgccaga agcagcttct cgacatgcac ttcaagcgct atcacgaccc caacttcgtc
     2281 cctgcggctt ttgtctgttc taagtgtggg aaaacattta cacgtcggaa taccatggca
     2341 agacatgctg ataattgtgc tggcccagat ggcgtagagg gggaaaatgg aggagaaacg
     2401 aagaagagta aacgtggaag aaaaagaaag atgcgctcta agaaagaaga ttcctctgac
     2461 agtgaaaatg ctgaaccaga tctggacgac aatgaggatg aggaggagcc tgccgtagaa
     2521 attgaacctg agccagagcc tcagcctgtg accccagccc caccacccgc caagaagcgg
     2581 agaggacgac cccctggcag aaccaaccag cccaaacaga accagccaac agctatcatt
     2641 caggttgaag accagaatac aggtgcaatt gagaacatta tagttgaagt aaaaaaagag
     2701 ccagatgctg agcccgcaga gggagaggaa gaggaggccc agccagctgc cacagatgcc
     2761 cccaacggag acctcacgcc cgagatgatc ctcagcatga tggaccggcc ggatccaccg
     2821 gtcatggtga gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg
     2881 aatggcgatg tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc
     2941 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc
     3001 accctggtga ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg
     3061 aagcagcacg acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc
     3121 ttcttcgagg atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc
     3181 ctggtgaatc gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc
     3241 aataagatgg agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag
     3301 aatggcatca aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg
     3361 gccgaccact accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac
     3421 cactacctgt ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg
     3481 atctacttcg gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag
     3541 tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3601 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3661 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3781 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3841 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3901 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3961 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4021 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4081 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4141 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4201 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4261 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4381 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4501 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4561 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4621 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4681 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4741 tcgatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4801 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4861 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4921 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     4981 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     5041 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     5101 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     5161 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     5221 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     5281 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     5341 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     5401 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5461 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5521 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5581 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5641 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     5701 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5761 caaaggaata gagttctaga ggatcataat cagccatacc acatttgtag aggttttact
     5821 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     5881 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5941 tttcacaaat aaagcatttt tttcactgcc tcgcgatcca caaccatggc gaaagggcct
     6001 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6181 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6241 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6301 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6361 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6421 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6481 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6541 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6601 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6661 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6721 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6781 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6841 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6901 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6961 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7021 aggtgcctca ctgattaagc attggtaagg cgcgccaggc cctgccatag cctcaggtta
     7081 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7141 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7201 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7261 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7321 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7381 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7441 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7501 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7561 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7621 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7681 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7741 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7801 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7861 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7921 tattaccgcc atgcat
//
