LOCUS       sequence-176632-        7984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan color variant of bright luminescent protein, fused to 
            b-tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7984)
  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 7984)
  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: tubulin-CeNL_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        511..714
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     665..685
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     759..778
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        759..777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2199..2201,2202..2204,2205..2717)
                     /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(2244..2265)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2852..2873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3433..3451)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3434..3451)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3471..3488)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3477..3701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3747..4175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3834..3853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4044..4065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4184..4204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4189..4518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4369..4504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4431..4450
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4585..5379
                     /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(4639..4658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5249..5268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5553..5674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5590..5609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5644..5663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5723..5739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5723..5739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5736..5758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5747..5763
                     /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(5771..5777,5778..5795,5796..5801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5816..5837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5954..5971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6125..6713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6205..6224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6884..7675,7676..7744))
                     /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     7507..7526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7745..7849)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7924..7943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
       61 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      121 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      181 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      241 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      301 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      361 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      421 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      481 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      541 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      601 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      661 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      721 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      781 acccaagctt cgccaccatg agggaaatcg tgcacatcca ggccggacag tgtggcaacc
      841 agatcggtgc taagttctgg gaggtgataa gcgatgaaca tggcatcgac cccaccggta
      901 cctaccacgg tgacagcgac ctgcagctgg accgaatctc tgtgtactat aatgaagcca
      961 caggtggcaa gtatgtccct cgagctatct tggtggatct agaacctggg actatggact
     1021 ccgttcgctc aggtcctttt ggccagatct tcagaccaga caacttcgtt ttcggtcagt
     1081 ctggggcagg caacaactgg gctaaaggcc actacacaga gggagctgag ttggttgact
     1141 ctgtcttgga tgtggtgcgg aaggaggcgg agagctgtga ttgcctgcaa ggctttcagc
     1201 tgacccactc actgggtgga ggcactggct ctggcatggg caccctgctc atcagcaaga
     1261 tccgggaaga atatcctgac cgtatcatga ataccttcag tgtggtgccc tcgcccaaag
     1321 tgtctgatac cgtggtcgag ccctacaatg ccaccctgtc tgtccatcag ttggttgaga
     1381 acacggatga gacctactgc atcgacaacg aggccctcta cgacatctgc ttccgtaccc
     1441 tcaagctcac cacgccaacc tacggagacc tgaaccatct cgtctcggcc accatgagcg
     1501 gcgtcaccac ctgcctccgt ttcccgggcc agcttaatgc tgaccttcga aagctggctg
     1561 tcaacatggt gccattccca cgtctccact tcttcatgcc tggctttgcc cctctcacca
     1621 gccgtggaag ccagcagtac cgggccctca ctgtgcctga acttacccag caggtcttcg
     1681 atgccaagaa catgatggcc gcctgcgacc cgcgccacgg ccggtacctc acagttgccg
     1741 ccgtcttccg tggacggatg tccatgaagg aggtggatga gcagatgctc aacgtgcaga
     1801 acaagaatag cagctacttc gtggaatgga tccccaacaa tgtcaagaca gctgtctgtg
     1861 acatcccacc gcgtggcctc aagatggcag tcaccttcat tggaaacagc acagccatcc
     1921 aggagctctt caagcgcatc tctgagcagt ttacggctat gttccgccgg aaggctttcc
     1981 tccactggta tacgggtgag ggcatggacg agatggagtt caccgaagct gagagcaaca
     2041 tgaacgacct cgtctctgag taccagcagt accaggatgc caccgcggaa gaggaagagg
     2101 atttcggaga ggaggcagaa gaggaggccc agtcgacggt accgcgggcc cgggatccag
     2161 gatctggcgg aggaagtggg ggaggatctg gagaattcat ggtgagcaag ggcgaggagc
     2221 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2281 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2341 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgtcctggg
     2401 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2461 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2521 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2581 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacttta
     2641 gcgacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     2701 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     2761 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     2821 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2881 ccgggttgca tacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     2941 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     3001 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     3061 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     3121 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     3181 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     3241 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     3301 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     3361 tgaccggctg gcggctgtgc gaacgcattc tggcgtaagc ggccgctcga gcatgcatct
     3421 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3481 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3541 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3601 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3661 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3721 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3781 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3841 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3901 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3961 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4021 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4081 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4141 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4201 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4261 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4321 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4381 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4441 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4501 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4561 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4621 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4681 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4741 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4801 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4861 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4921 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4981 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5041 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5101 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5161 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5221 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5281 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5341 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5401 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5461 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5521 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5641 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5701 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5761 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5821 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5881 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5941 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6001 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6061 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7021 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7861 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7921 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7981 ggag
//
