LOCUS       sequence-176630-        7797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan color variant of bright luminescent protein, fused to actin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7797)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 7797)
  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 
            Commun."; date: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: CeNL-Actin_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     695..715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     789..808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        789..807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(828..830,831..833,834..1346)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(873..894)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1134..1153)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1481..1502
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3276..3294)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3277..3294)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3314..3331)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3320..3544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3590..4018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3677..3696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3887..3908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4027..4047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4032..4361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4212..4347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4274..4293
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4428..5222
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4482..4501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5092..5111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5396..5517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5433..5452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5487..5506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5566..5582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5566..5582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5579..5601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5590..5606
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5614..5620,5621..5638,5639..5644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5659..5680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5797..5814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5968..6556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6048..6067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6727..7518,7519..7587))
                     /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     7350..7369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7588..7692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7767..7786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
       61 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      121 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      781 atcgaaatta atacgactca ctatagggag acccaagctt cgacaccatg gtgagcaagg
      841 gcgaagagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      901 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      961 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1021 tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1081 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1141 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1201 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1261 actactttag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1321 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1381 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1441 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1501 tgaccgccgc cgggttgcat acactcgaag atttcgttgg ggactggcga cagacagccg
     1561 gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg
     1621 gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg
     1681 acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa
     1741 aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg
     1801 gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg
     1861 aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca
     1921 acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca
     1981 tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcgggtgga tcaggcggta
     2041 ccggcagcgg tggcggtggc agcggtggcg gtggcagcgg tggatccatg gatgatgata
     2101 tcgccgcgct cgtcgtcgac aacggctccg gcatgtgcaa ggccggcttc gcgggcgacg
     2161 atgccccccg ggccgtcttc ccctccatcg tggggcgccc caggcaccag ggcgtgatgg
     2221 tgggcatggg tcagaaggat tcctatgtgg gcgacgaggc ccagagcaag agaggcatcc
     2281 tcaccctgaa gtaccccatc gagcacggca tcgtcaccaa ctgggacgac atggagaaaa
     2341 tctggcacca caccttctac aatgagctgc gtgtggctcc cgaggagcac cccgtgctgc
     2401 tgaccgaggc ccccctgaac cccaaggcca accgcgagaa gatgacccag atcatgtttg
     2461 agaccttcaa caccccagcc atgtacgttg ctatccaggc tgtgctatcc ctgtacgcct
     2521 ctggccgtac cactggcatc gtgatggact ccggtgacgg ggtcacccac actgtgccca
     2581 tctacgaggg gtatgccctc ccccatgcca tcctgcgtct ggacctggct ggccgggacc
     2641 tgactgacta cctcatgaag atcctcaccg agcgcggcta cagcttcacc accacggccg
     2701 agcgggaaat cgtgcgtgac attaaggaga agctgtgcta cgtcgccctg gacttcgagc
     2761 aagagatggc cacggctgct tccagctcct ccctggagaa gagctacgag ctgcctgacg
     2821 gccaggtcat caccattggc aatgagcggt tccgctgccc tgaggcactc ttccagcctt
     2881 ccttcctggg catggagtcc tgtggcatcc acgaaactac cttcaactcc atcatgaagt
     2941 gtgacgtgga catccgcaaa gacctgtacg ccaacacagt gctgtctggc ggcaccacca
     3001 tgtaccctgg cattgccgac aggatgcaga aggagatcac tgccctggca cccagcacaa
     3061 tgaagatcaa gatcattgct cctcctgagc gcaagtactc cgtgtggatc ggcggctcca
     3121 tcctggcctc gctgtccacc ttccagcaga tgtggatcag caagcaggag tatgacgagt
     3181 ccggcccctc catcgtccac cgcaaatgct tctaggaatt ctgcagatat ccatcacact
     3241 ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     3301 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3361 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3421 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3481 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3541 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3601 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3661 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3721 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3781 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3841 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3901 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3961 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4021 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4141 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4201 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4261 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4321 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4381 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4441 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4501 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4561 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4621 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4681 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4741 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4801 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4861 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4921 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4981 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5041 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5101 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5161 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5221 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5281 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5341 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5401 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5461 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5521 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5581 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5641 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5701 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5761 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5821 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7741 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagc
//
