LOCUS       sequence-267776-        6682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the [N-terminus of human CTCF (a.a. 2-265)]-GFP in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6682)
  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 6682)
  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-Nterm-N1
FEATURES             Location/Qualifiers
     source          1..6682
                     /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             1570..2289
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2411..2532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2502..2521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2539..2994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2676..2697)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2888..2907
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3021..3125
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3127..3484
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3127..3318
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3150..3170)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3335..3470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3492..4517
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3492..4517
                     /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    4634..4715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4743..4761)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4829..4933
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4934..5002,5003..5794)
                     /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(5152..5171)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5980..6568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6469..6488
                     /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 caggcgtgaa gaagaccggc
     1441 agcagcggcg gtaccagcgg caagaagagc aaacgcgctg gcccagatgg cgtagagggg
     1501 gaaaatggag gagaaacgaa gaagagtaaa cgtggaagaa aaagaaagat gcgcggggat
     1561 ccaccggtca tggtgagcaa gggcgccgag ctgttcaccg gcatcgtgcc catcctgatc
     1621 gagctgaatg gcgatgtgaa tggccacaag ttcagcgtga gcggcgaggg cgagggcgat
     1681 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcctgtgccc
     1741 tggcccaccc tggtgaccac cctgagctac ggcgtgcagt gcttctcacg ctaccccgat
     1801 cacatgaagc agcacgactt cttcaagagc gccatgcctg agggctacat ccaggagcgc
     1861 accatcttct tcgaggatga cggcaactac aagtcgcgcg ccgaggtgaa gttcgagggc
     1921 gataccctgg tgaatcgcat cgagctgacc ggcaccgatt tcaaggagga tggcaacatc
     1981 ctgggcaata agatggagta caactacaac gcccacaatg tgtacatcat gaccgacaag
     2041 gccaagaatg gcatcaaggt gaacttcaag atccgccaca acatcgagga tggcagcgtg
     2101 cagctggccg accactacca gcagaatacc cccatcggcg atggccctgt gctgctgccc
     2161 gataaccact acctgtccac ccagagcgcc ctgtccaagg accccaacga gaagcgcgat
     2221 cacatgatct acttcggctt cgtgaccgcc gccgccatca cccacggcat ggatgagctg
     2281 tacaagtgag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2341 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2401 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2461 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2521 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2581 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2641 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2701 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2761 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2821 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2881 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2941 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3001 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3061 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3121 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3241 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3301 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3361 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3421 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3481 caaagatcga tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     3541 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     3601 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     3661 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     3721 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     3781 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     3841 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     3901 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     3961 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     4021 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     4081 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     4141 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     4201 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     4261 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     4321 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     4381 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     4441 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     4501 cgagggcaaa ggaatagagt tctagaggat cataatcagc cataccacat ttgtagaggt
     4561 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4621 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4681 cacaaatttc acaaataaag catttttttc actgcctcgc gatccacaac catggcgaaa
     4741 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4801 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4861 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4921 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4981 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5041 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5101 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5161 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     5221 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5281 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5341 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5401 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5461 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5521 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5581 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5641 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5701 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5761 tgagataggt gcctcactga ttaagcattg gtaaggcgcg ccaggccctg ccatagcctc
     5821 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6661 taaccgtatt accgccatgc at
//
