LOCUS       sequence-12444-e        5476 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 5476)
  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 5476)
  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: C32V
FEATURES             Location/Qualifiers
     source          1..5476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(7..9,10..720)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             754..774
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(820..822,823..1536)
                     /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    1659..1780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1696..1715)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1750..1769
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1787..2242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1924..1945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2136..2155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2269..2373
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2375..2732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2375..2566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2398..2418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2583..2718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2767..3561
                     /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             2767..3561
                     /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(2821..2840)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3431..3450
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3749..3768)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3793..3840
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4169..4757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4658..4677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4932..5235
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5236..5439
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5390..5410
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5468..5476,1)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gatcctgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
       61 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      121 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      181 accctcgtga ccaccctgac ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg
      241 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      301 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      361 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      421 cacaagctgg agtacaacgc catcagcgac aacgtctata tcaccgccga caagcagaag
      481 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      541 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      601 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
      661 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtataag
      721 actagtggcc tcgagacgcg tgatatcaga tctgagaacc tgtacttcca gggcccgcgg
      781 gaattccccg ggggtaccgc tggaccggtc gccaccgtgg tgagcaaggg cgaggagctg
      841 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      901 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctcatc
      961 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct cggctacggc
     1021 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1081 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1141 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1201 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1261 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     1321 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     1381 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     1441 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1501 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     1561 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1621 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1681 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1741 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     1801 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1861 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1921 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1981 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2041 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2101 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2161 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2221 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2281 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2341 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2401 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2461 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2521 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2581 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2641 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2701 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2761 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2821 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2881 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2941 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3001 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3061 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3121 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3181 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3241 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3301 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3361 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3421 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3481 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3541 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3601 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3661 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3721 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3781 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3841 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3901 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3961 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4021 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4081 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4801 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4861 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     4921 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4981 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5041 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5101 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5161 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     5221 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     5281 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     5341 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     5401 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     5461 cgctagcgcc accatg
//
