LOCUS       sequence-267777-        7054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the N and C terminus of human CTCF-GFP with the zinc 
            finger domain removed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  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 7054)
  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-CTCFdZnF-N1
FEATURES             Location/Qualifiers
     source          1..7054
                     /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             1942..2661
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2783..2904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2874..2893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2911..3366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3048..3069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3260..3279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3393..3497
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3499..3856
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3499..3690
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3522..3542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3707..3842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3864..4889
                     /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
                     GDSQYKVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5006..5087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5115..5133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5201..5305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5306..5374,5375..6166)
                     /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(5524..5543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6352..6940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6841..6860
                     /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 gcgctctaag
     1561 aaagaagatt cctctgacag tgaaaatgct gaaccagatc tggacgacaa tgaggatgag
     1621 gaggagcctg ccgtagaaat tgaacctgag ccagagcctc agcctgtgac cccagcccca
     1681 ccacccgcca agaagcggag aggacgaccc cctggcagaa ccaaccagcc caaacagaac
     1741 cagccaacag ctatcattca ggttgaagac cagaatacag gtgcaattga gaacattata
     1801 gttgaagtaa aaaaagagcc agatgctgag cccgcagagg gagaggaaga ggaggcccag
     1861 ccagctgcca cagatgcccc caacggagac ctcacgcccg agatgatcct cagcatgatg
     1921 gaccggccgg atccaccggt catggtgagc aagggcgccg agctgttcac cggcatcgtg
     1981 cccatcctga tcgagctgaa tggcgatgtg aatggccaca agttcagcgt gagcggcgag
     2041 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2101 ctgcctgtgc cctggcccac cctggtgacc accctgagct acggcgtgca gtgcttctca
     2161 cgctaccccg atcacatgaa gcagcacgac ttcttcaaga gcgccatgcc tgagggctac
     2221 atccaggagc gcaccatctt cttcgaggat gacggcaact acaagtcgcg cgccgaggtg
     2281 aagttcgagg gcgataccct ggtgaatcgc atcgagctga ccggcaccga tttcaaggag
     2341 gatggcaaca tcctgggcaa taagatggag tacaactaca acgcccacaa tgtgtacatc
     2401 atgaccgaca aggccaagaa tggcatcaag gtgaacttca agatccgcca caacatcgag
     2461 gatggcagcg tgcagctggc cgaccactac cagcagaata cccccatcgg cgatggccct
     2521 gtgctgctgc ccgataacca ctacctgtcc acccagagcg ccctgtccaa ggaccccaac
     2581 gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg ccgccgccat cacccacggc
     2641 atggatgagc tgtacaagtg agcggccgcg actctagatc ataatcagcc ataccacatt
     2701 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2761 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2821 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2881 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2941 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3001 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3061 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3121 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3181 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3241 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3301 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3361 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3421 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3481 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3601 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3721 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3781 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3841 cctaggcttt tgcaaagatc gatatgaaaa agcctgaact caccgcgacg tctgtcgaga
     3901 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     3961 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     4021 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     4081 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     4141 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     4201 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     4261 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     4321 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     4381 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     4441 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     4501 cggtcattga ctggagcgag gcgatgttcg gggattccca atacaaggtc gccaacatct
     4561 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     4621 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     4681 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     4741 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     4801 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     4861 ccagcactcg tccgagggca aaggaataga gttctagagg atcataatca gccataccac
     4921 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4981 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcctc gcgatccaca
     5101 accatggcga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5161 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5221 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5281 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5341 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5401 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5461 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5521 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     5581 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5641 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5701 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5761 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5821 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5881 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5941 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6001 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6061 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6121 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaggcg cgccaggccc
     6181 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     6241 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6301 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6361 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6421 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6481 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6541 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6601 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6661 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6721 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6781 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6841 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6901 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6961 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7021 tgattctgtg gataaccgta ttaccgccat gcat
//
