LOCUS       sequence-102514-        4642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MXS_chaining vector with 2 cassettes .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4642)
  AUTHORS   Sladitschek HL, Neveu PA
  TITLE     MXS-Chaining: A Highly Efficient Cloning Platform for Imaging and 
            Flow Cytometry Approaches in Mammalian Systems.
  JOURNAL   PLoS One. 2015 Apr 24;10(4):e0124958. doi: 
            10.1371/journal.pone.0124958. eCollection 2015.
  PUBMED    25909630
REFERENCE   2  (bases 1 to 4642)
  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: "PLoS One.";
            date: "2015-04-24"; volume: "10(4):e0124958. doi"; pages: " 
            10.1371/journal.pone.0124958. eCollection 2015"
COMMENT     Sequence Label: EmptyTranscriptionalReporter
FEATURES             Location/Qualifiers
     source          1..4642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      124..133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             136..156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(163..165,166..168,169..879)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(208..229)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(469..488)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     816..837
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    898..1122
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1188..1491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1492..1695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1646..1666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1741..1750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1753..1773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1780..2484
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1922..1940)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2190..2209
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2503..2727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2750..2854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2855..2923,2924..3715)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3073..3092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3886..4474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4375..4394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgatatca aacgcgtaac tcgagtatgt cgaggtggcg tgtacggtgg gaggcctata
       61 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag aattcagatc tcagctggtc
      121 gaggccacca tggctcctaa gaagaagagg aaggtggtcg agatggtgag caagggcgag
      181 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      241 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      301 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
      361 tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      421 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      481 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      541 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      601 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      661 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      721 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
      781 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      841 gccgccggga tcactctcgg catggacgag ctgtacaaga cagatccatg agtcgagctg
      901 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
      961 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1021 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1081 aagacaatag caggcatgct ggggatgcgg tgggctctat ggtcgagtag ttattaatag
     1141 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     1201 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     1261 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     1321 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     1381 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1441 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1501 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1561 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1621 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1681 tatataagca gagctggttt agtgaaccgt cagatccgct agggatctca gctggtcgag
     1741 gccaccatgg ctcctaagaa gaagaggaag gtggtcgagg tgagcaaggg cgaggaggat
     1801 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     1861 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     1921 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     1981 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacattcc cgactacttg
     2041 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     2101 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     2161 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     2221 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     2281 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     2341 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     2401 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     2461 ggcggcatgg acgagctgta caagtctaca gattgagtcg agctgtgcct tctagttgcc
     2521 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2581 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2641 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2701 atgctgggga tgcggtgggc tctatggtcg accaattcgg ggaaatgtgc gcggaacccc
     2761 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2821 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2881 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2941 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3001 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3061 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3121 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3181 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3241 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3301 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3361 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3421 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3481 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3541 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3601 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3661 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3721 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3781 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3841 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3901 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3961 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4021 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4081 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4141 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4201 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4261 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4321 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4381 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4441 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4501 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4561 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4621 cgagcgcagc gagtcagtga gc
//
