LOCUS       sequence-478920-        7280 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses LATeNT* (slow reversing) in the mammalian cytosol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7280)
  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 7280)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 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-LATeNT(slow)
FEATURES             Location/Qualifiers
     source          1..7280
                     /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"
     promoter        complement(2833..2851)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2834..2851)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2871..2888)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2877..3101
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3147..3575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3234..3253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3444..3465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3584..3604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3589..3918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3769..3904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3831..3850
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3985..4779
                     /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(4039..4058)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4649..4668
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4953..5074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4990..5009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5044..5063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5123..5139)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5123..5139)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5136..5158)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5147..5163
                     /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(5171..5177,5178..5195,5196..5201))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5216..5237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5354..5371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5525..6113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5605..5624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6284..7075,7076..7144))
                     /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     6907..6926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7145..7249)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 caccggtccg atcaccatca acaacttccg ttactccgac
     1021 ccggttaaca acgacaccat catcatgatg gaaccgccgt actgcaaagg tctagacatc
     1081 tactacagag ccttcaaaat caccgaccgt atctggatcg ttccggaacg ttacgaattt
     1141 ggtaccaaac cggaagactt caacccgccg tcctccctga tcgaaggtgc ttccgaatac
     1201 tacgacccga actacctgcg taccgactcc gacaaagacc gtttcctgca gaccatggtt
     1261 aaactgttca accgtatcaa aaacaacgtt gctggtgaag ctctgctgga caaaatcatc
     1321 aacgctatcc cgtacctggg taactcctac tccctgctgg acaaattcga caccaactcc
     1381 aactccgttt ccttcaacct gctggaacag gatccgtccg gtgctaccac caaatccgct
     1441 atgctgacca acctgatcat cttcggtccg ggtccggttc tgaacaaaaa cgaagttcgt
     1501 ggtatcgttc tgcgtgttga caacaaaaac tacttcccgt gccgtgacgg tttcggttcc
     1561 atcatgcaga tggctttctg cccggaatac gtaccgacct tcgacaacgt tatcgaaaac
     1621 atcacctccc tgaccatcgg taaatccaaa tacttccagg acccggctct gctgctgatg
     1681 cacgaactga tccacgttct gcacggtctg tacggcatgc aggtttcctc ccacgaaatc
     1741 atcccgtcca aacaggaaat ctacatgcag cacacctacc cgatctccgc tgaagagctc
     1801 ttcaccttcg gtggtcagga cgctaacctg atctccatcg atatcaaaaa cgacctgtac
     1861 gaaaaaaccc tgaacgacta caaagctatc gctaacaaac tgtcccaggt tacctcctgc
     1921 aacgacccga acatcgacat cgactcctac aaacagatct atcagcagaa gtatcagttt
     1981 gataaggatt ccaacggcca gtatatagtg aacgaggata agtttcagat actgtataac
     2041 tccataatgt atggcgggag cggcagcggg gggtccggta gcggggagtt tcgggcaacc
     2101 acactggaac ggatcgagaa aagcttcctg attactgatc cgagactgcc tgacaaccca
     2161 atcatttttg tgagcaattc cttcctgcag ctgacagaat attctcggga agagatcctg
     2221 gggcgcaatt gccgttttct gcagggaccc gagacagacc gtgccactgt tcggaaaatc
     2281 agagatgcta ttgacaacca gactgaagtg accgttcagc tgatcaatta taccaagagc
     2341 ggcaagaagt tctggaacgt gttccacctg cagccgatgc gcgattataa gggcgacgtc
     2401 cagtacttca ttggcgtgca gctggatggc accgaacgcc ttcatggcgc cgctgagcgt
     2461 gaggcggtct gcctggtcaa aaagacagcc tttcagattt ttactgagat agagctgggc
     2521 aagaagttta acataaagac tcgcttaagc tatttttcca tgaaccacga tcctgtgaag
     2581 atacctaacc tgctggatga tactatatat aacgatactg agggctttaa catagagtcc
     2641 aaggatctga agtccgagta taagggccag aacatgaggg tgaacactaa cgcctttagg
     2701 aacgtcgacg gttccggtct ggtttccaaa ctgatcggtc tgtgcaaaaa aatcatcccg
     2761 ccgaccaaca tccgtgaaaa cctgtacaac cgtaccgctt aataactcga gcatgcatct
     2821 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     2881 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2941 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3001 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3061 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3121 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3481 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3541 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3601 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3721 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3841 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3901 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3961 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4021 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4081 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4141 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4201 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4261 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4321 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4381 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4441 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4501 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4561 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4621 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4681 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4741 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4801 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4861 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4921 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4981 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5041 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5101 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5161 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5221 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5761 gctccaagct gggctgtgtg cacaaacccc ccgttcagcc cgaccgctgc gccttatccg
     5821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5941 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6301 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6361 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6421 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6481 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6541 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6601 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6661 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6721 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6781 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6841 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6901 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6961 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7021 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7081 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7141 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7201 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7261 gaaaagtgcc acctgacgtc
//
