LOCUS       sequence-267778-        6669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the [N-terminus of human CTCF (a.a. 2-265)]-mBanana with a
            nuclear export signal in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6669)
  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 6669)
  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: pmBanana-Hyg-CTCF-NtermNES-N1
FEATURES             Location/Qualifiers
     source          1..6669
                     /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             1519..1545
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     regulatory      1561..1570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1567..2277
                     /codon_start=1
                     /product="yellow monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mBanana
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFCYGSKAYVKHPTGIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVAQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYSAETKTTYKAKKPVQLPGAYIAGEKIDITSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1712..1730)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1980..1999
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2398..2519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2489..2508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2526..2981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2663..2684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2875..2894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3008..3112
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3114..3471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3114..3305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3137..3157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3322..3457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3479..4504
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3479..4504
                     /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    4621..4702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4730..4748)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4816..4920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4921..4989,4990..5781)
                     /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(5139..5158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5967..6555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6456..6475
                     /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 gagaaacgct gccgccgctg gaacgcctga ccctgccgga tccaccggtc
     1561 gccaccatgg tgagcaaggg cgaggagaat aacatggccg tcatcaagga gttcatgcgc
     1621 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1681 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1741 ctgcccttcg cctgggacat cctgtcccct cagttctgtt acggctccaa ggcctacgtg
     1801 aagcacccca ctggtatccc cgactacttc aagctgtcct tccccgaggg cttcaagtgg
     1861 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tggctcagga ctcctccctg
     1921 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1981 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2041 gacggcgccc tgaagggcga gatcaagatg aggctgaagc tgaaggacgg cggccactac
     2101 agcgccgaga ccaagaccac ctacaaggcc aagaagcccg tgcagttgcc cggcgcctac
     2161 atagccggcg agaagatcga catcacctcc cacaatgagg actacactat cgtggaattg
     2221 tacgagcgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtagcgg
     2281 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2341 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2401 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2461 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2521 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3121 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatat
     3481 gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3541 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3601 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3661 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     3721 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3781 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     3841 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3901 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3961 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4021 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4081 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4141 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4201 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     4261 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4321 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4381 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     4441 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4501 atagagttct agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4561 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4621 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4681 aataaagcat ttttttcact gcctcgcgat ccacaaccat ggcgaaaggg cctcgtgata
     4741 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4801 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4861 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4921 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4981 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5041 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5101 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5161 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5221 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5281 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5341 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5401 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5461 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5521 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5581 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5641 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5701 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5761 tcactgatta agcattggta aggcgcgcca ggccctgcca tagcctcagg ttactcatat
     5821 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5881 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5941 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6001 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6061 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6121 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6181 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6241 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6301 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6361 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6421 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6481 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6541 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6601 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6661 gccatgcat
//
