LOCUS       sequence-266190-        6208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the [C-terminus of human CTCF (a.a. 577-728)]-GFP in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6208)
  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 6208)
  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-Cterm-N1
FEATURES             Location/Qualifiers
     source          1..6208
                     /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             1096..1815
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1937..2058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2028..2047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2065..2520)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2202..2223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2414..2433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2547..2651
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2653..3010
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2653..2844
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2676..2696)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2861..2996
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3018..4043
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3018..4043
                     /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    4160..4241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4269..4287)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4355..4459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4460..4528,4529..5320)
                     /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(4678..4697)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /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 gctggcccag atggcgtaga gggggaaaat
      661 ggaggagaaa cgaagaagag taaacgtgga agaaaaagaa agatgcgctc taagaaagaa
      721 gattcctctg acagtgaaaa tgctgaacca gatctggacg acaatgagga tgaggaggag
      781 cctgccgtag aaattgaacc tgagccagag cctcagcctg tgaccccagc cccaccaccc
      841 gccaagaagc ggagaggacg accccctggc agaaccaacc agcccaaaca gaaccagcca
      901 acagctatca ttcaggttga agaccagaat acaggtgcaa ttgagaacat tatagttgaa
      961 gtaaaaaaag agccagatgc tgagcccgca gagggagagg aagaggaggc ccagccagct
     1021 gccacagatg cccccaacgg agacctcacg cccgagatga tcctcagcat gatggaccgg
     1081 ccggatccac cggtcatggt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc
     1141 ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag
     1201 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct
     1261 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac
     1321 cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag
     1381 gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc
     1441 gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc
     1501 aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc
     1561 gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc
     1621 agcgtgcagc tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg
     1681 ctgcccgata accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag
     1741 cgcgatcaca tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat
     1801 gagctgtaca agtgagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     1861 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1921 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2041 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2101 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2161 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2221 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2281 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2341 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2401 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2461 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2521 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2581 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2641 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2701 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2761 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2821 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2881 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2941 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3001 cttttgcaaa gatcgatatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     3061 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     3121 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     3181 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     3241 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     3301 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     3361 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     3421 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     3481 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     3541 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     3601 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     3661 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     3721 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     3781 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     3841 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     3901 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     3961 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     4021 ctcgtccgag ggcaaaggaa tagagttcta gaggatcata atcagccata ccacatttgt
     4081 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4141 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4201 tagcatcaca aatttcacaa ataaagcatt tttttcactg cctcgcgatc cacaaccatg
     4261 gcgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4321 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4381 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4441 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4501 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4561 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4621 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4681 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4741 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4801 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4861 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4921 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4981 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5041 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5101 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5161 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5221 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5281 gatcgctgag ataggtgcct cactgattaa gcattggtaa ggcgcgccag gccctgccat
     5341 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6181 tgtggataac cgtattaccg ccatgcat
//
