LOCUS       sequence-404036-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Koushik SV, Chen H, Thaler C, Puhl HL, Vogel SS
  TITLE     Cerulean, Venus, and VenusY67C FRET reference standards.
  JOURNAL   Biophys J. 2006 Dec 15. 91(12):L99-L101.
  PUBMED    17040988
REFERENCE   2  (bases 1 to 5431)
  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: "Biophys J. 
            2006 Dec 15. 91(12):L99-L101."
COMMENT     Sequence Label: C5V
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(6..8,9..719)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    880..1001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(917..936)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     971..990
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1008..1463)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1145..1166)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1357..1376
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1490..1594
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1596..1953
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1596..1787
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1619..1639)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1804..1939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1988..2782
                     /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             1988..2782
                     /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(2042..2061)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2652..2671
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2970..2989)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3014..3061
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3390..3978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3879..3898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4153..4456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4457..4660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4611..4631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4699..4708
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4705..4707,4708..4710,4711..5421)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 gatctgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
       61 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      121 acggcaagct gaccctgaag ctcatctgca ccaccggcaa gctgcccgtg ccctggccca
      181 ccctcgtgac caccctcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
      241 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      301 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      361 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      421 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
      481 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
      541 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      601 actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
      661 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
      721 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata
      781 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      841 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      901 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
      961 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     1021 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1081 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     1141 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1201 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     1261 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1321 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1381 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1441 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1501 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1561 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     1621 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1681 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     1741 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     1801 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1861 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     1921 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     1981 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2041 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     2101 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     2161 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     2221 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     2281 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     2341 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     2401 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     2461 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     2521 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     2581 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     2641 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2701 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2761 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2821 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2881 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2941 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     3001 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     3061 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3121 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3181 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     3241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4081 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4141 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4201 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4261 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4321 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4381 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4441 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4501 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4561 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4621 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     4681 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     4741 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4801 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     4861 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
     4921 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4981 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     5041 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     5101 gaggacggca acatcctggg gcacaagctg gagtacaacg ccatcagcga caacgtctat
     5161 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     5221 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     5281 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     5341 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     5401 ggcatggacg agctgtacaa gtccggactc a
//
