LOCUS       sequence-437493-        6565 bp    DNA     circular SYN 15-APR-2025
DEFINITION  FLPo recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  TITLE     Plasmids to Generate AAVs for Non-Invasive SERT-Based Gap Junction 
            Tracing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6565)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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-CAG-FLPo-3xmiR122-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..792
                     /label=chicken beta-actin promoter
     primer_bind     1122..1144
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1206..1225
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1277..1286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1283..1288,1289..1309,1310..2581)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    2692..3280
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2745..2765)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3163..3174)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3312..3788
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3511..3530)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3828..3968
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3828..3957
                     /label=AAV2 ITR
     rep_origin      4043..4498
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4130..4149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4340..4361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4515..4534)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4634..4656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4694..4712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4780..4884
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4885..4953,4954..5745)
                     /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(5103..5122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5916..6504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6405..6424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg
      661 ggsgrggggs ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
     1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
     1261 cggtaccgga tccgccgcca ccatggctcc taagaagaag aggaaggtga tgagccagtt
     1321 cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga
     1381 gaggcccagc ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat
     1441 gatcacccac aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat
     1501 cagcaacagc ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca
     1561 gaaggccacc atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat
     1621 catcccttac aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca
     1681 gctgcagttc gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct
     1741 gaaggccctg ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag
     1801 cttcgagtac accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc
     1861 cacattcatc aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa
     1921 gctggtgcag aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac
     1981 aagcgtgtcc aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta
     2041 cctggacgag ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa
     2101 cagcagcagc aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa
     2161 caaggccctg aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag
     2221 ccacatcggc aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac
     2281 aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac
     2341 ccaccagatc accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta
     2401 cgaccccatc agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg
     2461 gcagcacatc gagcagctga agggcagcgc cgagggcagc atcagatacc ccgcctggaa
     2521 cggcatcatc agccaggagg tgctggacta cctgagcagc tacatcaaca ggcggatctg
     2581 agaattcacc ggtcaaacac cattgtcaca ctccatacta tcaaacacca ttgtcacact
     2641 ccagatcaac aaacaccatt gtcacactcc acgccggcga agcttatcga taatcaacct
     2701 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2761 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2821 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2881 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2941 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3001 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3061 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3121 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3181 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3241 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3301 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3361 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3421 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3481 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3541 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3601 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3661 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3721 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3781 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     3841 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3901 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3961 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4021 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4081 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4141 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4201 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4261 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4321 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4381 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4441 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4501 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4561 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4621 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4681 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4741 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4861 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4921 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4981 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5041 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5101 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5161 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5221 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5281 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5341 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5401 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5461 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5521 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5581 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5641 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5701 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5761 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5821 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5881 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5941 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6001 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6061 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6121 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6181 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6241 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6301 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6361 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6421 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6481 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6541 ttttgctggc cttttgctca catgt
//
