LOCUS       sequence-474922-        8397 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  Expresses Ca-LATeNT in the mammalian cytosol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8397)
  AUTHORS   Roh H, Kim D, Kim B, Jeon Y, Kim Y, Jacko M, Xu F, Lin C, Won Um J, 
            Ting AY
  TITLE     Light-activated tetanus neurotoxin for conditional proteolysis and 
            inducible synaptic inhibition in vivo
  JOURNAL   bioRxiv 2025.01.27.635161; doi: 10.1101/2025.01.27.635161
REFERENCE   2  (bases 1 to 8397)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.01.27.635161; doi: 10.1101/2025.01.27.635161"
COMMENT     Sequence Label: pcDNA3-Ca-LATeNT
FEATURES             Location/Qualifiers
     source          1..8397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             910..951
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1477..1533
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     promoter        complement(3949..3967)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3950..3967)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3987..4004)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3993..4217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4263..4691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4350..4369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4560..4581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4700..4720)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4705..5035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4886..5021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4948..4967
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5102..5896
                     /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(5156..5175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5766..5785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6070..6191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6107..6126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6161..6180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6240..6256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6240..6256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6253..6275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6264..6280
                     /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(6288..6294,6295..6312,6313..6318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6333..6354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6471..6488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6642..7230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6722..6741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7401..8192,8193..8261))
                     /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     8024..8043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8262..8366)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gccaccatgg gcaagcccat ccccaacccc ctgctgggcc tggacagcac caagggctcg
      961 ggctcgacct cgggctcggg caccggtttc acgttggaag attttgtcgg tgattgggag
     1021 caaaccgcgg cctacaacct cgatcaagtg cttgaacaag gcggagtatc cagtgttctg
     1081 caaaccctgg ccgtatctgt gactccgatt cagaggattg tgcgcagcgg cgaaaatgga
     1141 ctcaaaattg atattcacgt catcattcct tatgaggggc tgtcagccga tcaaatggca
     1201 cacatcgagg aagtatttaa agttgtttac cccgtggacg atcatcactt taaggtgatt
     1261 atggaatatg gcactcttgt aatagacggg gttacaccca acatgcttaa ttactttggg
     1321 agaccttatg agggtatagc agttttcgat ggcaagaaga ttaccgtgac aggcaccctt
     1381 tggaacggca acaaaataat cgacgagcgg ctcatcactc cggaaggaag tatgctcttc
     1441 cgtgtcacga ttaattctgc atccagcgat tcatcccggc gtaaatggaa caaaacaggt
     1501 catgccgtca gggccattgg acgcctgagc tcaggtggga gtggcggatc aggcggcagc
     1561 ggtggatccg gcggttccaa tttgcctgat caactgactg aggaacaaat tgccgagttc
     1621 aaggaagaat ttagtctgtt cgacaaagac ggggatggga ccattaccac caaggaatta
     1681 ggcaccgtca tgaggtcctt gggacaaaac cccacagaag ctgaacttca ggacatgatt
     1741 aatgaggttg atgcagatgg cgacggaact attgacttcc ctgaatttct gactatgatg
     1801 gcgaggaaaa tgaagtatcg cgacactgag gaagagatta gagaggcctt tggcgtcttt
     1861 gataaagacg ggatcggcta tataagcgca gcagagttga ggcatgtgat gacgaacctg
     1921 ggagagaaac ttaccgatga agaagtggat gaaatgatac gggaagcgga tattgatggc
     1981 gatggtcagg taaactatga agaatttgtg caaatgatga cagcgaaatc ccccgcggta
     2041 actggatacc ggcttctcga agaaatctca aacgggagcg gcagcggggg gtccggtagc
     2101 gggccgatca ccatcaacaa cttccgttac tccgacccgg ttaacaacga caccatcatc
     2161 atgatggaac cgccgtactg caaaggtcta gacatctact acagagcctt caaaatcacc
     2221 gaccgtatct ggatcgttcc ggaacgttac gaatttggta ccaaaccgga agacttcaac
     2281 ccgccgtcct ccctgatcga aggtgcttcc gaatactacg acccgaacta cctgcgtacc
     2341 gactccgaca aagaccgttt cctgcagacc atggttaaac tgttcaaccg tatcaaaaac
     2401 aacgttgctg gtgaagctct gctggacaaa atcatcaacg ctatcccgta cctgggtaac
     2461 tcctactccc tgctggacaa attcgacacc aactccaact ccgtttcctt caacctgctg
     2521 gaacaggatc cgtccggtgc taccaccaaa tccgctatgc tgaccaacct gatcatcttc
     2581 ggtccgggtc cggttctgaa caaaaacgaa gttcgtggta tcgttctgcg tgttgacaac
     2641 aaaaactact tcccgtgccg tgacggtttc ggttccatca tgcagatggc tttctgcccg
     2701 gaatacgtac cgaccttcga caacgttatc gaaaacatca cctccctgac catcggtaaa
     2761 tccaaatact tccaggaccc ggctctgctg ctgatgcacg aactgatcca cgttctgcac
     2821 ggtctgtacg gcatgcaggt ttcctcccac gaaatcatcc cgtccaaaca ggaaatctac
     2881 atgcagcaca cctacccgat ctccgctgaa gagctcttca ccttcggtgg tcaggacgct
     2941 aacctgatct ccatcgatat caaaaacgac ctgtacgaaa aaaccctgaa cgactacaaa
     3001 gctatcgcta acaaactgtc ccaggttacc tcctgcaacg acccgaacat cgacatcgac
     3061 tcctacaaac agatctatca gcagaagtat cagtttgata aggattccaa cggccagtat
     3121 atagtgaacg aggataagtt tcagatactg tataactcca taatgtatgg cgggagcggc
     3181 agcggggggt ccggtagcgg ggagtttcgg gcaaccacac tggaacggat cgagaaaagc
     3241 ttcgtgatta ctgatccgag actgcctgac aacccaatca tttttgtgag caattccttc
     3301 ctgcagctga cagaatattc tcgggaagag atcctggggc gcaattgccg ttttctgcag
     3361 ggacccgaga cagaccgtgc cactgttcgg aaaatcagag atgctattga caaccagact
     3421 gaagtgaccg ttcagctgat caattatacc aagagcggca agaagttctg gaacgtgttc
     3481 cacctgcagc cgatgcgcga ttataagggc gacgtccagt acttcattgg cgtgcagctg
     3541 gatggcaccg aacgccttca tggcgccgct gagcgtgagg cggtctgcct ggtcaaaaag
     3601 acagcctttc agatttttac tgagatagag ctgggcaaga agtttaacat aaagactcgc
     3661 ttaagctatt tttccatgaa ccacgatcct gtgaagatac ctaacctgct ggatgatact
     3721 atatataacg atactgaggg ctttaacata gagtccaagg atctgaagtc cgagtataag
     3781 ggccagaaca tgagggtgaa cactaacgcc tttaggaacg tcgacggttc cggtctggtt
     3841 tccaaactga tcggtctgtg caaaaaaatc atcccgccga ccaacatccg tgaaaacctg
     3901 tacaaccgta ccgcttaata actcgagcat gcatctagag ggccctattc tatagtgtca
     3961 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4021 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4081 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4141 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4201 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4261 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4321 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4381 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4441 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4501 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4561 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4621 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4681 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4741 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4861 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4921 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4981 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5041 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     5101 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5161 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5221 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5281 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5341 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5401 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5461 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5521 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5581 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5641 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5701 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5761 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5821 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5881 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5941 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6001 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6061 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6181 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6241 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6301 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6361 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6421 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7081 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
