LOCUS       sequence-267780-        6222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the [C-terminus of human CTCF (a.a. 577-728)]-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 6222)
  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 6222)
  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-CtermNES-N1
FEATURES             Location/Qualifiers
     source          1..6222
                     /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             1072..1098
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     regulatory      1114..1123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1120..1830
                     /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(1265..1283)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1533..1552
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1951..2072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2042..2061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2079..2534)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2216..2237)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2428..2447
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2561..2665
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2667..3024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2667..2858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2690..2710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2875..3010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3032..4057
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3032..4057
                     /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    4174..4255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4283..4301)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4369..4473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4474..4542,4543..5334)
                     /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(4692..4711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5520..6108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6009..6028
                     /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 tctaagaaag aagattcctc tgacagtgaa
      661 aatgctgaac cagatctgga cgacaatgag gatgaggagg agcctgccgt agaaattgaa
      721 cctgagccag agcctcagcc tgtgacccca gccccaccac ccgccaagaa gcggagagga
      781 cgaccccctg gcagaaccaa ccagcccaaa cagaaccagc caacagctat cattcaggtt
      841 gaagaccaga atacaggtgc aattgagaac attatagttg aagtaaaaaa agagccagat
      901 gctgagcccg cagagggaga ggaagaggag gcccagccag ctgccacaga tgcccccaac
      961 ggagacctca cgcccgagat gatcctcagc atgatggacc ggggtaccag cggcaagaag
     1021 agcaaacgcg ctggcccaga tggcgtagag ggggaaaatg gaggagaaac gctgccgccg
     1081 ctggaacgcc tgaccctgcc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     1141 aataacatgg ccgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg
     1201 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1261 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1321 cctcagttct gttacggctc caaggcctac gtgaagcacc ccactggtat ccccgactac
     1381 ttcaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1441 ggcgtggtga ccgtggctca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1501 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     1561 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     1621 atgaggctga agctgaagga cggcggccac tacagcgccg agaccaagac cacctacaag
     1681 gccaagaagc ccgtgcagtt gcccggcgcc tacatagccg gcgagaagat cgacatcacc
     1741 tcccacaatg aggactacac tatcgtggaa ttgtacgagc gcgccgaggg ccgccactcc
     1801 accggcggca tggacgagct gtacaagtag cggccgcgac tctagatcat aatcagccat
     1861 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1921 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2041 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     2101 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2161 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2221 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2281 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2341 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2401 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2461 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2521 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2641 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     2701 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2761 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2821 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2881 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2941 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3001 tttggaggcc taggcttttg caaagatcga tatgaaaaag cctgaactca ccgcgacgtc
     3061 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3121 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3181 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     3241 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
     3301 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3361 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     3421 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3481 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3541 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3601 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     3661 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     3721 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     3781 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     3841 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     3901 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     3961 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4021 ccgacgcccc agcactcgtc cgagggcaaa ggaatagagt tctagaggat cataatcagc
     4081 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4141 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4201 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcctcgc
     4261 gatccacaac catggcgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4321 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4681 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     4741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaggcgcg
     5341 ccaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5401 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5461 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5521 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5581 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5641 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5701 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5761 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5821 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5881 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5941 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6001 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6061 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6121 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6181 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
