LOCUS       sequence-275043-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytoskeleton, Excitation: 604, Emission: 659.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  AUTHORS   Chu J, Haynes RD, Corbel SY, Li P, Gonzalez-Gonzalez E, Burg JS, 
            Ataie NJ, Lam AJ, Cranfill PJ, Baird MA, Davidson MW, Ng HL, Garcia 
            KC, Contag CH, Shen K, Blau HM, Lin MZ
  TITLE     Non-invasive intravital imaging of cellular differentiation with a 
            bright red-excitable fluorescent protein.
  JOURNAL   Nat Methods. 2014 Mar 16. doi: 10.1038/nmeth.2888.
  PUBMED    24633408
REFERENCE   2  (bases 1 to 5957)
  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 
            Methods. 2014 Mar 16. doi: 10.1038/nmeth.2888."
COMMENT     Sequence Label: mCardinal-Myotilin-C-14
FEATURES             Location/Qualifiers
     source          1..5957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        899..1102
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1053..1073
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1141..1150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1147..1878
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    3544..3665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3581..3600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3635..3654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3672..4127)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3809..3830)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4021..4040
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4154..4258
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4283..4303)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4327..4537
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4349..4545
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4396..4531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4580..5374
                     /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             4580..5374
                     /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(4634..4653)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5244..5263
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5562..5581)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5606..5653
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      781 agttccgcgt tacataactt acggtaaatg gcccgcctgg cattatgccc agtacatgac
      841 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      901 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      961 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1021 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1081 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
     1141 gccaccatgg tgagcaaggg cgaggagctg atcaaggaga acatgcacat gaagctgtac
     1201 atggaaggca ccgtgaacaa ccaccacttc aagtgcacca ccgaagggga gggcaagccc
     1261 tacgagggca cccagaccca gaggattaag gtggtggagg gaggccccct gccgttcgca
     1321 ttcgacatcc tggccacctg ctttatgtac gggagcaaga ccttcatcaa ccacacccag
     1381 ggcatccccg atttctttaa gcagtccttc cctgagggct tcacatggga gagagtcacc
     1441 acatacgaag acgggggcgt gcttaccgtt acccaggaca ccagcctcca ggacggctgc
     1501 ttgatctaca acgtcaagct cagaggggtg aacttcccat ccaacggccc tgtgatgcag
     1561 aagaaaacac tcggctggga ggccaccacc gagaccctgt accccgctga cggcggcctg
     1621 gaaggcagat gcgacatggc cctgaagctc gtgggcgggg gccacctgca ctgcaacctg
     1681 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctacttt
     1741 gtggaccgca gactggaaag aatcaaggag gccgacaatg agacctacgt cgagcagcac
     1801 gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacaa acttaatggc
     1861 atggacgagc tgtacaagtc cggaagtgct gggagcgcaa gcggaagcgg aggctccggt
     1921 tttaactacg aacgtccaaa acacttcatc cagtcccaaa acccatgtgg ctccagattg
     1981 cagcctcctg gaccagaaac ctccagcttc tctagccaga ccaaacagtc ttccattatc
     2041 atccagcccc gccagtgtac agagcaaaga ttttctgcct cctcaacact gagctctcac
     2101 atcaccatgt cctcctctgc tttccctgct tctccccagc agcatgctgg ctccaaccca
     2161 ggccaaaggg ttacaaccac ctataaccag tccccagcca gcttcctcag ctccatatta
     2221 ccatcacagc ctgattacaa tagcagtaaa atcccttccg ctatggattc caactatcaa
     2281 cagtcctcag ctggccaacc tataaatgca aagccatccc aaactgcaaa tgctaagccc
     2341 ataccaagaa ctcctgatca tgaaatacaa ggatcaaaag aagctttgat tcaagatttg
     2401 gaaagaaagc tgaaatgcaa ggacaccctt cttcataatg gaaatcaacg tctaacatat
     2461 gaagagaaga tggctcgcag attgctagga ccacagaatg cagctgctgt gtttcaagct
     2521 caggatgaca gtggtgcaca agactcgcag caacacaact cagaacatgc gcgactgcaa
     2581 gttcctacat cacaagtaag aagtagatca acctcaaggg gagatgtgaa tgatcaggat
     2641 gcaatccagg agaaatttta cccaccacgt ttcattcaag tgccagagaa catgtcgatt
     2701 gatgaaggaa gattctgcag aatggacttc aaagtgagtg gactgccagc tcctgatgtg
     2761 tcatggtatc taaatggaag aacagttcaa tcagatgatt tgcacaaaat gatagtgtct
     2821 gagaagggtc ttcattcact catctttgaa gtagtcagag cttcagatgc aggggcttat
     2881 gcatgtgttg ccaagaatag agcaggagaa gccaccttca ctgtgcagct ggatgtcctt
     2941 gcaaaagaac ataaaagagc accaatgttt atctacaaac cacagagcaa aaaagtttta
     3001 gagggagatt cagtgaaact agaatgccag atctcggcta tacctccacc aaagcttttc
     3061 tggaaaagaa ataatgaaat ggtacaattc aacactgacc gaataagctt atatcaagat
     3121 aacactggaa gagttacttt actgataaaa gatgtaaaca agaaagatgc tgggtggtat
     3181 actgtgtcag cagttaatga agctggagtg actacatgta acacaagatt agacgttacg
     3241 gcacgtccaa accaaactct tccagctcct aagcagttac gggttcgacc aacattcagc
     3301 aaatatttag cacttaatgg gaaaggtttg aatgtaaaac aagcttttaa cccagaagga
     3361 gaatttcagc gtttggcagc tcaatctgga ctctatgaaa gtgaagaact ttaaggatcc
     3421 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3481 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3541 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3601 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3661 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3721 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3781 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3841 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3901 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3961 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4021 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4081 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4141 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4201 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4261 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4321 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4561 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4621 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4681 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4741 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4801 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4861 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4921 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4981 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5041 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5101 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5161 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5221 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5281 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5341 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5401 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5461 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5521 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5581 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5641 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5701 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5761 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5821 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5941 cactgagcgt cagaccc
//
