LOCUS       sequence-127742-        6085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  One half component of the optimized Cre recombinase dependent on GFP
            (CRE-DOG OPT) for rAAV production and expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6085)
  AUTHORS   Tang JC, Rudolph S, Dhande OS, Abraira VE, Choi S, Lapan SW, Drew 
            IR, Drokhlyansky E, Huberman AD, Regehr WG, Cepko CL
  TITLE     Cell type-specific manipulation with GFP-dependent Cre recombinase.
  JOURNAL   Nat Neurosci. 2015 Sep;18(9):1334-41. doi: 10.1038/nn.4081. Epub 
            2015 Aug 10.
  PUBMED    26258682
REFERENCE   2  (bases 1 to 6085)
  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"; doi: "10.1038/nn.4081"; 
            journalName: "Nat Neurosci"; date: "2015-09"; volume: "18"; issue: 
            "9"; pages: "1334-41"
COMMENT     Sequence Label: pAAV-EF1a-N-CretrcintG
FEATURES             Location/Qualifiers
     source          1..6085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1432..1441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1468..1488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     gap             1795..2184
                     /estimated_length=390
     misc_feature    2212..2800
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2265..2285)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2683..2694)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2832..3308
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3031..3050)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3348..3488
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3348..3477
                     /label=AAV2 ITR
     rep_origin      3563..4018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3650..3669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3860..3881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4035..4054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4154..4176
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4214..4232)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4300..4404
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4405..4473,4474..5265)
                     /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(4623..4642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5436..6024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5925..5944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg
     1441 gataaagcgg aattaattcc cgagcctcca aaaaagaaga gaaaggtcga attgggtacc
     1501 acctctgatg aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct
     1561 gaacacacct ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg
     1621 aacctcgtcg aaagctatcg caaggccagc aacaaggcct actttgagtg gaccatcgaa
     1681 gccagggacc tctccctgct gggaagccac gtgcgcaagg ccacctacca gacctacgcc
     1741 cccatcctgt acgctagcgg aggaggtggt ggaggtggag gtggaggtca attgnnnnnn
     1801 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1861 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1921 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1981 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     2041 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     2101 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     2161 nnnnnnnnnn nnnnnnnnnn nnnntaagaa ttcgatatca agcttatcga taatcaacct
     2221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2581 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     2641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2761 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     2821 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     2881 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2941 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3001 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3061 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3121 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3181 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3241 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3301 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     3361 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3421 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3481 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3541 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3601 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3661 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3721 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     3781 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3841 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3901 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3961 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4021 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4081 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4141 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4201 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4261 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4321 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4381 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4441 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4501 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4561 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4621 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4681 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4741 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4801 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4861 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4921 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4981 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5041 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5101 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5161 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5221 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5281 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5341 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5401 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5461 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5521 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5581 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5641 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5701 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5761 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5821 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5881 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5941 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6001 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6061 ttttgctggc cttttgctca catgt
//
