LOCUS       sequence-363598-        7145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the nuclear localized YFP-CCTd(peptide of 
            Cav1.2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  AUTHORS   Yang Y, Yu Z, Geng J, Liu M, Liu N, Li P, Hong W, Yue S, Jiang H, Ge
            H, Qian F, Xiong W, Wang P, Song S, Li X, Fan Y, Liu X
  TITLE     Cytosolic peptides encoding Ca(V)1 C-termini downregulate the 
            calcium channel activity-neuritogenesis coupling.
  JOURNAL   Commun Biol. 2022 May 19;5(1):484. doi: 10.1038/s42003-022-03438-1.
  PUBMED    35589958
REFERENCE   2  (bases 1 to 7145)
  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: "Commun 
            Biol."; date: "2022-05-19"; volume: "5(1):484. doi"; pages: " 
            10.1038/s42003-022-03438-1"
COMMENT     Sequence Label: pcDNA3-NLS-YFP-CCTc
FEATURES             Location/Qualifiers
     source          1..7145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(386..1177,1178..1246))
                     /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     1009..1028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1247..1351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1426..1445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1617..1996
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1997..2200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2151..2171
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2245..2264
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2245..2263
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2295..2315
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2316..2318,2319..2321,2322..2834)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(2361..2382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2622..2641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2969..2990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4080..4098)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4081..4098)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4118..4135)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4124..4348
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4394..4822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4481..4500)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4691..4712
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4831..4851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4836..5165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5016..5151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5078..5097
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5232..6026
                     /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(5286..5305)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5896..5915
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6200..6321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6237..6256)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6291..6310
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6370..6386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6370..6386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6383..6405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6394..6410
                     /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(6418..6424,6425..6442,6443..6448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6463..6484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6601..6618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6772..7145,1..215))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6852..6871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
       61 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      241 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      361 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      421 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      481 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      541 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      601 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      661 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
      721 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      781 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      841 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
      901 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
      961 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1021 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1081 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1141 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1201 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1381 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     1441 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     1501 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     1561 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     1621 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1681 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1741 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1801 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1861 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1921 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1981 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2041 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2101 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2161 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     2221 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
     2281 ccgccaccat gccaccaaag aagaagcgca aggtgatggt gagcaagggc gaggagctgt
     2341 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2401 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2461 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcg
     2521 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2581 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2641 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2701 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2761 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2821 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2881 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
     2941 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3001 ggatcactct cgcggccgct gagggccacg ggcccccctt gtcccctgcc atccgggtgc
     3061 aggaggtggc gtggaagctc agctccaaca ggtgccactc ccgggagagc caggcagcca
     3121 tggcgggtca ggaggagacg tctcaggatg agacctatga agtgaagatg aaccatgaca
     3181 cggaggcctg cagtgagccc agcctgctct ccacagagat gctctcctac caggatgacg
     3241 aaaatcggca actgacgctc ccagaggagg acaagaggga catccggcaa tctccgaaga
     3301 ggggtttcct ccgctctgcc tcactaggtc gaagggcctc cttccacctg gaatgtctga
     3361 agcgacagaa ggaccgaggg ggagacatct ctcagaagac agtcctgccc ttgcatctgg
     3421 ttcatcatca ggcattggca gtggcaggcc tgagccccct cctccagaga agccattccc
     3481 ctgcctcatt ccctaggcct tttgccaccc caccagccac acctggcagc cgaggctggc
     3541 ccccacagcc cgtccccacc ctgcggcttg agggggtcga gtccagtgag aaactcaaca
     3601 gcagcttccc atccatccac tgcggctcct gggctgagac cacccccggt ggcgggggca
     3661 gcagcgccgc ccggagagtc cggcccgtct ccctcatggt gcccagccag gctggggccc
     3721 cagggaggca gttccacggc agtgccagca gcctggtgga agcggtcttg atttcagaag
     3781 gactggggca gtttgctcaa gatcccaagt tcatcgaggt caccacccag gagctggccg
     3841 acgcctgcga catgaccata gaggagatgg agagcgcggc cgacaacatc ctcagcgggg
     3901 gcgccccaca gagccccaat ggcgccctct taccctttgt gaactgcagg gacgcggggc
     3961 aggaccgagc cgggggcgaa gaggacgcgg gctgtgtgcg cgcgcggggt cgaccgagtg
     4021 aggaggagct ccaggacagc agggtctacg tcagcagcct gtaatctaga gggccctatt
     4081 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     4141 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4201 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4261 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4321 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4381 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4441 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4501 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4561 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4621 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4681 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4741 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4801 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4861 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4981 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5041 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5101 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5161 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5221 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5281 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5341 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5401 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5461 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5521 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5581 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5641 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5701 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5761 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5821 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5881 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5941 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6001 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6061 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6121 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6181 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6241 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6301 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6361 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6601 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7141 tagca
//
