LOCUS       sequence-363599-        7289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the cytosolic localized YFP-CCTd(peptide of 
            Cav1.3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7289)
  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 7289)
  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-NES-YFP-CCTd
FEATURES             Location/Qualifiers
     source          1..7289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1211..1230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1402..1781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1782..1985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1936..1956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2030..2049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2030..2048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2113..2115,2116..2118,2119..2631)
                     /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(2158..2179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2419..2438)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2766..2787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4009..4027)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4010..4027)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4047..4064)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4053..4277
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4323..4751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4410..4429)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4620..4641
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4760..4780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4765..5094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4945..5080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5007..5026
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5161..5955
                     /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(5215..5234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5825..5844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6129..6250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6166..6185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6220..6239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6299..6315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6299..6315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6312..6334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6323..6339
                     /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(6347..6353,6354..6371,6372..6377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6392..6413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6530..6547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6701..7289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6781..6800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     1381 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     1441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1741 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1981 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     2041 actataggga gacccaagct tggtaccgcc accatgcttg ctcttaagtt ggctggactt
     2101 gatattggtt ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2161 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2221 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2281 ccctggccca ccctcgtgac caccttcggc tacggcgtgc agtgcttcgc ccgctacccc
     2341 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2401 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2461 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2521 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2581 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2641 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2701 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     2761 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgc ggccgctaat
     2821 atgtccaaag ctgcccatgg aaagcggccc agcattggga acctcgagca tgtgtctgaa
     2881 aatgggcatc attcttccca caagcatgac cgggagcctc agagaaggtc cagtgtgaaa
     2941 aggtccgact caggagatga acagctccca actatttgcc gggaagaccc agagatacat
     3001 ggctatttca gggaccccca ctgcttgggg gagcaggagt atttcagtag tgaggaatgc
     3061 tacgaggatg acagctcgcc cacctggagc aggcaaaact atggctacta cagcagatac
     3121 ccaggcagaa acatcgactc tgagaggccc cgaggctacc atcatcccca aggattcttg
     3181 gaggacgatg actcgcccgt ttgctatgat tcacggagat ctccaaggag acgcctacta
     3241 cctcccaccc cagcatccca ccggagatcc tccttcaact ttgagtgcct gcgccggcag
     3301 agcagccagg aagaggtccc gtcgtctccc atcttccccc atcgcacggc cctgcctctg
     3361 catctaatgc agcaacagat catggcagtt gccggcctag attcaagtaa agcccagaag
     3421 tactcaccga gtcactcgac ccggtcgtgg gccacccctc cagcaacccc tccctaccgg
     3481 gactggacac cgtgctacac ccccctgatc caagtggagc agtcagaggc cctggaccag
     3541 gtgaacggca gcctgccgtc cctgcaccgc agctcctggt acacagacga gcccgacatc
     3601 tcctaccgga ctttcacacc agccagcctg actgtcccca gcagcttccg gaacaaaaac
     3661 agcgacaagc agaggagtgc ggacagcttg gtggaggcag tcctgatatc cgaaggcttg
     3721 ggacgctatg caagggaccc aaaatttgtg tcagcaacaa aacacgaaat cgctgatgcc
     3781 tgtgacctca ccatcgacga gatggagagt gcagccagca ccctgcttaa tgggaacgtg
     3841 cgtccccgag ccaacgggga tgtgggcccc ctctcacacc ggcaggacta tgagctacag
     3901 gactttggtc ctggctacag cgacgaagag ccagaccctg ggagggatga ggaggacctg
     3961 gcggatgaaa tgatatgcat caccaccttg taatctagag ggccctattc tatagtgtca
     4021 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4081 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4141 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4201 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4261 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4321 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4741 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5101 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5341 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5941 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6001 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6061 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6121 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6241 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7261 agattacgcg cagaaaaaaa ggatctcaa
//
