LOCUS       sequence-479181-        7257 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Expresses LATeNT in mammalian cells; CaMKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7257)
  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 7257)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 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: pAAV-CaMKII-LATeNT
FEATURES             Location/Qualifiers
     source          1..7257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        799..2087
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      2108..2117
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2117..2158
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4034..4622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4087..4107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4505..4516)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4654..5130
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4853..4872)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5170..5310
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5170..5299
                     /label=AAV2 ITR
     rep_origin      5385..5840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5472..5491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5682..5703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5857..5876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5976..5998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6036..6054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6122..6226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6227..6295,6296..7087)
                     /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     complement(6445..6464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg ggttcttctt
      841 ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc tgcccagggt
      901 cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa attccatgcc
      961 ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag atgaccaatg
     1021 agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca gccctgggtg
     1081 agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt gacgggaagc
     1141 acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc ctctgggagg
     1201 atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca gtggtgacag
     1261 ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt ttggtacttc
     1321 ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg agagctggaa
     1381 agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag gaagcaaaga
     1441 aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt cccctgtaaa
     1501 ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca gaggagatga
     1561 agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg tggcacccac
     1621 tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt ctgttaaaag
     1681 ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg gactaagttt
     1741 gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag ggtccacttg
     1801 ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa agaggagcag
     1861 gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa cgggaacagg
     1921 gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc gaggctgtga
     1981 gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt tctccgtttg
     2041 cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct agagcggtac
     2101 cggatccgcc accatgggca agcccatccc caaccccctg ctgggcctgg acagcaccaa
     2161 gggctcgggc tcgacctcgg gctcgggcac cggtccgatc accatcaaca acttccgtta
     2221 ctccgacccg gttaacaacg acaccatcat catgatggaa ccgccgtact gcaaaggtct
     2281 agacatctac tacagagcct tcaaaatcac cgaccgtatc tggatcgttc cggaacgtta
     2341 cgaatttggt accaaaccgg aagacttcaa cccgccgtcc tccctgatcg aaggtgcttc
     2401 cgaatactac gacccgaact acctgcgtac cgactccgac aaagaccgtt tcctgcagac
     2461 catggttaaa ctgttcaacc gtatcaaaaa caacgttgct ggtgaagctc tgctggacaa
     2521 aatcatcaac gctatcccgt acctgggtaa ctcctactcc ctgctggaca aattcgacac
     2581 caactccaac tccgtttcct tcaacctgct ggaacaggat ccgtccggtg ctaccaccaa
     2641 atccgctatg ctgaccaacc tgatcatctt cggtccgggt ccggttctga acaaaaacga
     2701 agttcgtggt atcgttctgc gtgttgacaa caaaaactac ttcccgtgcc gtgacggttt
     2761 cggttccatc atgcagatgg ctttctgccc ggaatacgta ccgaccttcg acaacgttat
     2821 cgaaaacatc acctccctga ccatcggtaa atccaaatac ttccaggacc cggctctgct
     2881 gctgatgcac gaactgatcc acgttctgca cggtctgtac ggcatgcagg tttcctccca
     2941 cgaaatcatc ccgtccaaac aggaaatcta catgcagcac acctacccga tctccgctga
     3001 agagctcttc accttcggtg gtcaggacgc taacctgatc tccatcgata tcaaaaacga
     3061 cctgtacgaa aaaaccctga acgactacaa agctatcgct aacaaactgt cccaggttac
     3121 ctcctgcaac gacccgaaca tcgacatcga ctcctacaaa cagatctatc agcagaagta
     3181 tcagtttgat aaggattcca acggccagta tatagtgaac gaggataagt ttcagatact
     3241 gtataactcc ataatgtatg gcgggagcgg cagcgggggg tccggtagcg gggagtttcg
     3301 ggcaaccaca ctggaacgga tcgagaaaag cttcgtgatt actgatccga gactgcctga
     3361 caacccaatc atttttgtga gcaattcctt cctgcagctg acagaatatt ctcgggaaga
     3421 gatcctgggg cgcaattgcc gttttctgca gggacccgag acagaccgtg ccactgttcg
     3481 gaaaatcaga gatgctattg acaaccagac tgaagtgacc gttcagctga tcaattatac
     3541 caagagcggc aagaagttct ggaacgtgtt ccacctgcag ccgatgcgcg attataaggg
     3601 cgacgtccag tacttcattg gcgtgcagct ggatggcacc gaacgccttc atggcgccgc
     3661 tgagcgtgag gcggtctgcc tggtcaaaaa gacagccttt cagattttta ctgagataga
     3721 gctgggcaag aagtttaaca taaagactcg cttaagctat ttttccatga accacgatcc
     3781 tgtgaagata cctaacctgc tggatgatac tatatataac gatactgagg gctttaacat
     3841 agagtccaag gatctgaagt ccgagtataa gggccagaac atgagggtga acactaacgc
     3901 ctttaggaac gtcgacggtt ccggtctggt ttccaaactg atcggtctgt gcaaaaaaat
     3961 catcccgccg accaacatcc gtgaaaacct gtacaaccgt accgcttgag aattcgatat
     4021 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4081 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4141 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4201 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4261 cgcaaccccc actggttggg gaattgccac cacctgtcag ctcctttccg gaactttcgc
     4321 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4381 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     4441 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4501 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4561 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4621 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     4681 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4741 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4801 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4861 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4921 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4981 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     5041 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     5101 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     5161 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     5221 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     5281 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     5341 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     5401 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5461 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5521 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5581 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5641 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5701 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5761 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5821 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5881 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5941 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6001 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6061 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6121 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6181 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6241 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6301 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6361 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6421 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6481 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6541 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6601 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6661 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6721 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6781 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6841 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6901 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6961 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7021 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7081 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7141 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7201 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
