LOCUS       sequence-264399-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HcRed-NPM1c (cytoplasmic mutant, human) in mammalian cells
            through transient transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  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 5571)
  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: pHcRed-NPM1c-C1
FEATURES             Location/Qualifiers
     source          1..5571
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1296
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     polyA_signal    2359..2480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2396..2415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2450..2469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2487..2942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2624..2645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2836..2855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2969..3073
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3075..3432
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3075..3266
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3098..3118)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3283..3418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3467..4261
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3467..4261
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3521..3540)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4131..4150
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4449..4468)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4493..4540
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4869..5457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5358..5377
                     /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 ccggtcgcca ccatggtgag cggcctgctg aaggagagta tgcgcatcaa gatgtacatg
      661 gagggcaccg tgaacggcca ctacttcaag tgcgagggcg agggcgacgg caaccccttc
      721 gccggcaccc agagcatgag aatccacgtg accgagggcg cccccctgcc cttcgccttc
      781 gacatcctgg ccccctgctg cgagtacggc agcaggacct tcgtgcacca caccgccgag
      841 atccccgact tcttcaagca gagcttcccc gagggcttca cctgggagag aaccaccacc
      901 tacgaggacg gcggcatcct gaccgcccac caggacacca gcctggaggg caactgcctg
      961 atctacaagg tgaaggtgca cggcaccaac ttccccgccg acggccccgt gatgaagaac
     1021 aagagcggcg gctgggagcc cagcaccgag gtggtgtacc ccgagaacgg cgtgctgtgc
     1081 ggccggaacg tgatggccct gaaggtgggc gaccggcacc tgatctgcca ccactacacc
     1141 agctaccgga gcaagaaggc cgtgcgcgcc ctgaccatgc ccggcttcca cttcaccgac
     1201 atccggctcc agatgctgcg gaagaagaag gacgagtact tcgagctgta cgaggccagc
     1261 gtggcccggt acagcgacct gcccgagaag gccaactccg gactcagatc tcgagctcaa
     1321 gcttcgaatt ctatgaaaga ttcgatggac atggacatga gccccctgag gccccagaac
     1381 tatcttttcg gttgtgaact aaaggccgac aaagattatc actttaaggt ggataatgat
     1441 gaaaatgagc accagttatc tttaagaacg gtcagtttag gggctggtgc aaaggatgag
     1501 ttgcacattg ttgaagcaga ggcaatgaat tacgaaggca gtccaattaa agtaacactg
     1561 gcaactttga aaatgtctgt acagccaacg gtttcccttg ggggctttga aataacacca
     1621 ccagtggtct taaggttgaa gtgtggttca gggccagtgc atattagtgg acagcactta
     1681 gtagctgtgg aggaagatgc agagtcagaa gatgaagagg aggaggatgt gaaactctta
     1741 agtatatctg gaaagcggtc tgcccctgga ggtggtagca aggttccaca gaaaaaagta
     1801 aaacttgctg ctgatgaaga tgatgacgat gatgatgaag aggatgatga tgaagatgat
     1861 gatgatgatg attttgatga tgaggaagct gaagaaaaag cgccagtgaa gaaatctata
     1921 cgagatactc cagccaaaaa tgcacaaaag tcaaatcaga atggaaaaga ctcaaaacca
     1981 tcatcaacac caagatcaaa aggacaagaa tccttcaaga aacaggaaaa aactcctaaa
     2041 acaccaaaag gacctagttc tgtagaagac attaaagcaa aaatgcaagc aagtatagaa
     2101 aaaggtggtt ctcttcccaa agtggaagcc aaattcatca attatgtgaa gaattgcttc
     2161 cggatgactg accaagaggc tattcaagat ctctgtctgg cagtggagga agtctcttta
     2221 agaaaatagg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     2281 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2341 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2401 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2461 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2521 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2581 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2641 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2701 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2761 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2821 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2881 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2941 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3001 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3061 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3361 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3421 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3481 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3541 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3601 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3661 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3721 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3781 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3841 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3901 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3961 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4021 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4081 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4141 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4201 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4261 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4321 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4381 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4441 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4501 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4561 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4621 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4681 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4741 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4801 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4981 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5221 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5281 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5341 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5401 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5461 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5521 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
