LOCUS       sequence-192715-        6627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent Cerulean-E2A-SADB19 rabies glycoprotein expression, 
            under hSyn promoter, for producing AAV virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6627)
  AUTHORS   none
  TITLE     SADB19 rabies virus tracing plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6627)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV hSyn-Flex RG-cerulean
FEATURES             Location/Qualifiers
     source          1..6627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      355..364
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    367..400
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    463..496
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2087..2146)
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             complement(join(2162..2872,2873..2875))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2204..2225)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2553..2572
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2812..2833
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2884..2917)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2988..3021)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3110..3586
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3309..3328)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3626..3766
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3626..3755
                     /label=AAV2 ITR
     rep_origin      3841..4296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3928..3947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4138..4159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4313..4332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4432..4454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4492..4510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4578..4682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4683..4751,4752..5543)
                     /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(4901..4920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5714..6302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6203..6222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6364..6493
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(6520..6627,1..340)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
       61 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
      121 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
      181 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
      241 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
      301 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggtcgccacc
      361 atggccataa cttcgtataa agtatcctat acgaagttat atcaaaatag gaagaccaat
      421 gcttcaccat cgacccgaat tgccaagcat caccatcgac ccataacttc gtataatgta
      481 tgctatacga agttatactt caactagttc acagtctggt ctcgccgccg ctcttgtggc
      541 tctcccagct gctgatgatc ttgccgctct ggggggtcac gctcacctct ctgccggtgc
      601 ctctcaggtt gtgctgggtg ggctcgcttc tgttcactct tctgcagcag gtcatcagga
      661 agatgatcag catcagggcg gtcagggcgc cggcgctcag cagcacgtac ttgccccagt
      721 tgggcaggcc caggtccacg ccgctcacct ggttgtgcac gtcgggcagg tgcacctcca
      781 cgaagtcctc ggcctcgtcg ccgtccttga acacggtgct ggggtcggcc agggggtgca
      841 ccagggggat cacgctgctc tccagcagct ccatgtgctg ctgcagcagg ctgctctgca
      901 tctcggggat cagcacgttg ccgtcggggc ccaggatgat gccgttgaag aacacgccgt
      961 tcacgtgggg gtggcatctg ccgcccactc tcaggcagcc cttgctgggc aggatctcgt
     1021 tccaggttct cacgctcttg tagtgggcgt cggcctccat cagggtcttg ttgaagatgg
     1081 tgtaggcctt gccgaagccg ggcaccagct ttctcaggtg gctcagtctt ctgaagctca
     1141 cgctcttggt ggtcatgatg ctctccaggg cgtccaggca ctcctctctc tttctcacca
     1201 gctcctccac caccaggtgc tcgatctcgt cgcttctgaa gtcgtgcagg ttcaccagct
     1261 tgtcgggggg gcaccacttg gtctcgttgc tggtctgcat gctcacccag gtgccgtcca
     1321 tcagtctcag gcccagcacg ccgcacagct tcagcttgca ggcgcccttc aggctcttgt
     1381 acaggcctct ctcgtccacg aagccgcagg tctcgctgcc cttgctggct ctcttgcctc
     1441 tgctgttggt gaagatgtcg cagctcatgc ccagtctggg gttctcgggc atccagatgg
     1501 tgtagtcgtg gttggtgctg cagtaggtgc tgctcacggc cacgccgctg cacttgccgc
     1561 tggggaacac tctgctgtgc aggcttctgt cgtaggggtc caggtcggcc acgctggggc
     1621 tgatgatcac caggctctcc ttggtggtct tcacggttct cagccatctg tagtcggggt
     1681 aggggttgtg caggctctcc tcgtatctgg ggtcgccggc catcttccag ttgtaggcgg
     1741 ctctgcaggc gtcgggggtg ggtctgaagt gctttctctt gaaggtggtg gtcacgtagc
     1801 ccacgaagtt ggtgtaggtc tcggcctcgg tcaccacgcc ggtgcaggtg aagccgttca
     1861 ccttgatggc caggatgtag cccaccttca gctccatgta gctgaagccg ctcaggttgg
     1921 tgcagccctc gtcctccacc accaggttgt tggggcagct caggtggtgg atgtcgatgg
     1981 ggctccaggg gcccagcttg tcggggatgg tgtagatggg gaacttgccg aagcacaggg
     2041 ggaacaccag caggggcacg aacagcaggg cctggggcac tccggaagga ccggggttac
     2101 tttcaacatc gccagcgagt ttcaacaaag cgtagttagt acattgacca ctaccgctag
     2161 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2221 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt
     2281 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     2341 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     2401 tcttctgctt gtcggcggtg atatagacgt tgtcgctgat ggcgttgtac tccagcttgt
     2461 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     2521 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     2581 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     2641 tcatgtggtc ggggtagcgg gcgaagcact gcacgcccca ggtcagggtg gtcacgaggg
     2701 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     2761 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     2821 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcacctcga
     2881 cgcataactt cgtataggat actttatacg aagttatctt tgggattctt cctattttga
     2941 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata
     3001 gcatacatta tacgaagtta tgtccctcga agaggttcga attccccggg gatcctctag
     3061 agtcgacctg cagaagcttg cctcgagcag cgctgctcga gagatctacg ggtggcatcc
     3121 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3181 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3241 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3301 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3361 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3421 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3481 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3541 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3601 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3661 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3721 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3781 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3841 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3901 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3961 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4021 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4081 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4141 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4201 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4261 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4321 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4381 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4441 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4501 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4561 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4681 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4741 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4801 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4861 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4921 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4981 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5041 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5101 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5161 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5221 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5281 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5341 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5401 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5461 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5521 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5581 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5641 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6361 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     6421 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     6481 actaggggtt cctgcggccg cacgcgggcc ctgcgtatga gtgcaagtgg gttttaggac
     6541 caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca
     6601 cccaaccccc attccccaaa ttgcgca
//
