LOCUS       sequence-264382-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HcRed-NPM1wildtype (human) in mammalian cells through 
            transient transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  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 5567)
  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-NPM1wt-C1
FEATURES             Location/Qualifiers
     source          1..5567
                     /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    2355..2476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2392..2411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2446..2465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2483..2938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2620..2641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2832..2851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2965..3069
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3071..3428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3071..3262
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3094..3114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3279..3414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3463..4257
                     /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             3463..4257
                     /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(3517..3536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4127..4146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4445..4464)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4489..4536
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4865..5453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5354..5373
                     /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 ctatggaaga 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 ctctggcagt ggaggaagtc tctttaagaa
     2221 aatagggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2281 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2341 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2401 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2461 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2521 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2581 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2641 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2701 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2761 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2821 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2881 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2941 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3001 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3061 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3361 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3421 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3481 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3541 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3601 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3661 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3721 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3781 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3841 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3901 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3961 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4021 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4081 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4141 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4201 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4261 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4321 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4381 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4441 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4501 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4561 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4621 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4681 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4741 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4801 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4861 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4921 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4981 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5041 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5101 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5161 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5221 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5281 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5341 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5401 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5461 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5521 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
