LOCUS       sequence-311208-        5648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry from LAP1_2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Maturana CJ, Verpeut JL, Pisano TJ, Dhanerawala ZM, Esteves A, 
            Enquist LW, Engel EA
  TITLE     Small Alphaherpesvirus Latency-Associated Promoters Drive Efficient 
            and Long-Term Transgene Expression in the CNS.
  JOURNAL   Mol Ther Methods Clin Dev. 2020 Apr 14;17:843-857. doi: 
            10.1016/j.omtm.2020.04.004. eCollection 2020 Jun 12.
  PUBMED    32368565
REFERENCE   2  (bases 1 to 5648)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2020-04-14"; volume: "17:843-857. doi"; 
            pages: " 10.1016/j.omtm.2020.04.004. eCollection 2020 Jun 12"
COMMENT     Sequence Label: pAAV-LAP1_2-mCherry
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             956..1666
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1101..1119)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1369..1388
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1691..2279
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1744..1764)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2162..2173)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2311..2787
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2510..2529)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2827..2967
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2827..2956
                     /label=AAV2 ITR
     rep_origin      3042..3497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3129..3148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3339..3360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3514..3533)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3633..3655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3693..3711)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3779..3883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3884..3952,3953..4744)
                     /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(4102..4121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4915..5503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5404..5423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5565..5648,1..46)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
       61 cccgggggag tcggggggtc ccagttcggc gcggtctcca cagcgcacaa agtccggctg
      121 cagtaaattt actgcgggtg cagttaccct ggcggccaca ccggcccgat ggcgctgcag
      181 taactctact gcagctacag tttactacag ttgcagtacc gcgcgccgcc gccaaatact
      241 acagtagatt tcctgtggcc gccgcgtact gcagtttacc gcggctgcag taaactgcag
      301 tatcgcgcgg taaattgcag tctggcagcc gcgcgttact gcaattagcg gtggctcccg
      361 acactctggc caattggtgc taatgggccg tgatggtcca tgtgggggtg atgtaaccgc
      421 cgggcccggt tgggcactca gatggtggcc gggcgccagg ccaaagtgct gtctgagtgc
      481 cactttatga ctttgttttt ctcaaacaac atcaattatg gatgcacatc gtgtatataa
      541 tccccggtcc gcgctccgcc cacccatcac agcagccgcg gacgctgcgc gccggagcgg
      601 tccatctcgc cagccagcca accagccgag ccgcccagcc gacccgagag ccccgagagc
      661 cagactccct cagccataga agacaccggg cgggagagac ggactgaaaa aatatatctt
      721 tttttatttt gtctgggcct ggagacccgc agcaggagcg gaggtgggtg cggggccggg
      781 agccggagca ggaccgggaa caggaacagg aacaggaaca ggaacaggaa caggagtggg
      841 gccgggagca ggagcaggag cgggagccga agtgggggca ggagcggcgg cggccgcagc
      901 agcaacaggg tcgccccagt ccgcggcgag gaagagggag ctcggatccg ccaccatggt
      961 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     1021 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1081 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1141 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     1201 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1261 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1321 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1381 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1441 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1501 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1561 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1621 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagaat tcgatatcaa
     1681 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1741 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1801 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1861 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1921 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     1981 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2041 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2101 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2161 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2221 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2281 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     2341 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     2401 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     2461 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     2521 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     2581 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     2641 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     2701 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     2761 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     2821 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2881 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2941 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3001 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3061 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3121 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3181 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3241 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3301 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3361 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3421 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3481 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3541 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3601 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3661 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3721 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3781 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3841 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3901 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3961 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4021 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4081 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4141 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4201 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4261 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4321 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4381 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4441 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4501 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4561 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4621 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4681 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4741 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4801 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4861 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4921 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4981 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5041 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5101 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5161 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5221 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5281 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5341 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5401 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5461 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5521 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     5581 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     5641 tgagcgag
//
