LOCUS       sequence-44976-e        5099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the nucleus (can be used for colocalization
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5099)
  AUTHORS   Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen 
            L, Hink MA, van Weeren L, Gadella TW Jr, Royant A
  TITLE     Structure-guided evolution of cyan fluorescent proteins towards a 
            quantum yield of 93%.
  JOURNAL   Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738.
  PUBMED    22434194
REFERENCE   2  (bases 1 to 5099)
  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"; journalName: "Nat 
            Commun."; date: "2012-03-20"; volume: "3:751. doi"; pages: " 
            10.1038/ncomms1738"
COMMENT     Sequence Label: pmTurquoise2-H2A
FEATURES             Location/Qualifiers
     source          1..5099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    565..686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(602..621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     656..675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(693..1148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(830..851)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1042..1061
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1175..1279
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1281..1638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1281..1472
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1304..1324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1489..1624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1673..2467
                     /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             1673..2467
                     /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(1727..1746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2337..2356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2655..2674)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2699..2746
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3075..3663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3564..3583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3838..4141
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4142..4345
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4296..4316
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4384..4393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4390..4392,4393..4395,4396..5099,1..7)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4435..4456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4696..4715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5043..5064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gtacaagtac tcagatctcg agctcaagct tcgaattctg atgtcgggac gcggcaagca
       61 gggaggcaaa gctcgcgcca aagccaagac ccgctcttct cgtgccggtc tccagttccc
      121 cgtgggccga gtgcaccgac tgctccgcaa gggcaactat gctgagcggg tcggggccgg
      181 cgcgccggtg tacctggcgg cggtgctgga gtacctgact gccgagatcc tggagctggc
      241 gggcaacgcc gcccgcgaca acaagaagac ccgcattatc ccgcgccact tgcagctggc
      301 catccgcaac gacgaggagc tcaacaagct gctgggcaaa gtaaccatcg ctcagggtgg
      361 tgtcctgccc aacatccagg ctgtgctact gcccaagaag accgagagtc accacaaggc
      421 caaaggcaaa taacgggatc caccggatct agataactga tcataatcag ccataccaca
      481 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
      541 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
      601 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      661 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
      721 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
      781 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
      841 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
      901 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
      961 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1021 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     1081 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     1141 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1201 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1261 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     1321 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1381 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1441 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1561 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1621 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     1681 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1741 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1801 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     1861 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1921 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1981 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2041 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2101 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2161 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     2221 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2281 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2341 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2401 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2461 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2521 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2581 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     2641 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2701 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     2761 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     2821 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     2881 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     2941 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3001 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3061 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3121 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3181 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3241 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3301 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3361 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3421 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3481 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3541 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3601 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3661 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3721 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     3781 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     3841 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3901 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3961 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4021 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4081 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4141 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4201 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4261 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4321 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     4381 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4441 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4501 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4561 tggcccaccc tcgtgaccac cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac
     4621 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4681 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4741 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4801 ctggggcaca agctggagta caactacttt agcgacaacg tctatatcac cgccgacaag
     4861 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     4921 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4981 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     5041 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagct
//
