LOCUS       sequence-266191-        6772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the [Zinc finger domain of human CTCF (a.a. 265-603)]-GFP 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6772)
  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 6772)
  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-ZnF-N1
FEATURES             Location/Qualifiers
     source          1..6772
                     /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             1660..2379
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2501..2622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2592..2611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2629..3084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2978..2997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3111..3215
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3217..3574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3217..3408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3240..3260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3425..3560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3582..4607
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3582..4607
                     /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    4724..4805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4833..4851)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4919..5023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5024..5092,5093..5884)
                     /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(5242..5261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6070..6658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6559..6578
                     /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 ttccagtgtg agctttgcag ttacacgtgt
      661 ccacggcgtt caaatttgga tcgtcacatg aaaagccaca ctgatgagag accacacaag
      721 tgccatctct gtggcagggc attcagaaca gtcaccctcc tgaggaatca ccttaacaca
      781 cacacaggta ctcgtcctca caagtgccca gactgcgaca tggcctttgt gaccagtgga
      841 gaattggttc ggcatcgtcg ttacaaacac acccacgaga agccattcaa gtgttccatg
      901 tgcgattacg ccagtgtaga agtcagcaaa ttaaaacgtc acattcgctc tcatactgga
      961 gagcgtccgt ttcagtgcag tttgtgcagt tatgccagca gggacacata caagctgaaa
     1021 aggcacatga gaacccattc aggggaaaag ccttatgaat gttatatttg tcatgctcgg
     1081 tttacccaaa gtggtaccat gaagatgcac attttacaga agcacacaga aaatgtggcc
     1141 aaatttcact gtccccactg tgacacagtc atagcccgaa aaagtgattt gggtgtccac
     1201 ttgcgaaagc agcattccta tattgagcaa ggcaagaaat gccgttactg tgatgctgtg
     1261 tttcatgagc gctatgccct catccagcat cagaagtcac acaagaatga gaagcgcttt
     1321 aagtgtgacc agtgtgatta cgcttgtaga caggagaggc acatgatcat gcacaagcgc
     1381 acccacaccg gggagaagcc ttacgcctgc agccactgcg ataagacctt ccgccagaag
     1441 cagcttctcg acatgcactt caagcgctat cacgacccca acttcgtccc tgcggctttt
     1501 gtctgttcta agtgtgggaa aacatttaca cgtcggaata ccatggcaag acatgctgat
     1561 aattgtgctg gcccagatgg cgtagagggg gaaaatggag gagaaacgaa gaagagtaaa
     1621 cgtggaagaa aaagaaagat gcgcggggat ccaccggtca tggtgagcaa gggcgccgag
     1681 ctgttcaccg gcatcgtgcc catcctgatc gagctgaatg gcgatgtgaa tggccacaag
     1741 ttcagcgtga gcggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1801 atctgcacca ccggcaagct gcctgtgccc tggcccaccc tggtgaccac cctgagctac
     1861 ggcgtgcagt gcttctcacg ctaccccgat cacatgaagc agcacgactt cttcaagagc
     1921 gccatgcctg agggctacat ccaggagcgc accatcttct tcgaggatga cggcaactac
     1981 aagtcgcgcg ccgaggtgaa gttcgagggc gataccctgg tgaatcgcat cgagctgacc
     2041 ggcaccgatt tcaaggagga tggcaacatc ctgggcaata agatggagta caactacaac
     2101 gcccacaatg tgtacatcat gaccgacaag gccaagaatg gcatcaaggt gaacttcaag
     2161 atccgccaca acatcgagga tggcagcgtg cagctggccg accactacca gcagaatacc
     2221 cccatcggcg atggccctgt gctgctgccc gataaccact acctgtccac ccagagcgcc
     2281 ctgtccaagg accccaacga gaagcgcgat cacatgatct acttcggctt cgtgaccgcc
     2341 gccgccatca cccacggcat ggatgagctg tacaagtgag cggccgcgac tctagatcat
     2401 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2461 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2521 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2581 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2641 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2701 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2761 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2821 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2881 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2941 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3001 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3061 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3121 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3181 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3481 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3541 aggaggcttt tttggaggcc taggcttttg caaagatcga tatgaaaaag cctgaactca
     3601 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     3661 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     3721 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     3781 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     3841 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     3901 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     3961 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     4021 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     4081 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     4141 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     4201 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     4261 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
     4321 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     4381 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     4441 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     4501 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     4561 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagagt tctagaggat
     4621 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4681 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4741 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4801 actgcctcgc gatccacaac catggcgaaa gggcctcgtg atacgcctat ttttataggt
     4861 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5041 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5161 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5221 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     5281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5401 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5581 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5641 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5881 gtaaggcgcg ccaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6061 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6181 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6661 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6721 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
