LOCUS       sequence-323618-        4746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: C1 Cloning Vector, Excitation: 604, Emission: 659.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4746)
  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 4746)
  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-C1
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      562..571
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             568..1299
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     misc_feature    1300..1365
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1489..1610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1526..1545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1580..1599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1617..2072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1754..1775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1966..1985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2099..2203
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2205..2562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2205..2396
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2228..2248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2413..2548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2597..3391
                     /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             2597..3391
                     /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(2651..2670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3261..3280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3579..3598)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3623..3670
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3999..4587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4488..4507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gatcaaggag
      601 aacatgcaca tgaagctgta catggaaggc accgtgaaca accaccactt caagtgcacc
      661 accgaagggg agggcaagcc ctacgagggc acccagaccc agaggattaa ggtggtggag
      721 ggaggccccc tgccgttcgc attcgacatc ctggccacct gctttatgta cgggagcaag
      781 accttcatca accacaccca gggcatcccc gatttcttta agcagtcctt ccctgagggc
      841 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgcttaccgt tacccaggac
      901 accagcctcc aggacggctg cttgatctac aacgtcaagc tcagaggggt gaacttccca
      961 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccaccac cgagaccctg
     1021 taccccgctg acggcggcct ggaaggcaga tgcgacatgg ccctgaagct cgtgggcggg
     1081 ggccacctgc actgcaacct gaagaccaca tacagatcca agaaacccgc taagaacctc
     1141 aagatgcccg gcgtctactt tgtggaccgc agactggaaa gaatcaagga ggccgacaat
     1201 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa
     1261 ctggggcaca aacttaatgg catggacgag ctgtacaagt ccggactcag atctcgagct
     1321 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa
     1381 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1441 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1501 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1561 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     1621 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1681 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1741 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1801 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1861 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1921 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1981 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2041 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2101 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2161 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2221 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2281 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2341 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2401 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2461 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2521 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2581 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2641 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2701 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2761 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2821 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2881 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2941 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3001 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3061 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3121 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3181 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3241 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3301 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3361 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3421 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3481 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3541 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3601 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     3661 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3721 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3781 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     3841 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4141 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4381 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4621 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4681 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     4741 tagccc
//
