LOCUS       sequence-127747-        6784 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 6784)
  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 6784)
  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-C-CreintG
FEATURES             Location/Qualifiers
     source          1..6784
                     /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"
     gap             1438..1827
                     /estimated_length=390
     misc_feature    2911..3499
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2964..2984)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3382..3393)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3531..4007
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3730..3749)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4047..4187
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4047..4176
                     /label=AAV2 ITR
     rep_origin      4262..4717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4349..4368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4559..4580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4734..4753)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4853..4875
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4913..4931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5104..5172,5173..5964)
                     /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(5322..5341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6135..6723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6624..6643
                     /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 cgccaccnnn
     1441 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1501 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1561 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1621 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1681 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1741 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1801 nnnnnnnnnn nnnnnnnnnn nnnnnnngct agcgtcctgc tgaatgtcct gagcaagtgc
     1861 gccggttcca aaaagttccg accagctcct gctgctgctt ttgctcgcga gtgccgcgga
     1921 ttctacttcg agctgcagga gctcaaggag gacgactact acggaatcac cctgtccgac
     1981 gattccgacc accaatttct gctggccaac caggtggtgg tccataattg taacaggaaa
     2041 tggttccctg ctgaacctga ggatgtgagg gactacctcc tgtacctgca agccagaggc
     2101 ctggctgtga agaccatcca acagcacctg ggccagctca acatgctgca caggagatct
     2161 ggcctgcctc gcccttctga ctccaatgct gtgtccctgg tgatgaggag aatcagaaag
     2221 gagaatgtgg atgctgggga gagagccaag caggccctgg cctttgaacg cactgacttt
     2281 gaccaagtca gatccctgat ggagaactct gacagatgcc aggacatcag gaacctggcc
     2341 ttcctgggca ttgcctacaa caccctgctg cgcattgccg aaattgccag aatcagagtg
     2401 aaggacatct cccgcaccga tggtgggaga atgctgatcc acattggcag gaccaagacc
     2461 ctggtgtcca cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa gctggtggag
     2521 agatggatct ctgtgtctgg tgtggctgat gaccccaaca actacctgtt ctgccgggtc
     2581 agaaagaatg gtgtggctgc cccttctgcc acctcccaac tgtccacccg ggccctggaa
     2641 gggatctttg aggccaccca ccgcctgatc tatggtgcca aggatgactc tgggcagaga
     2701 tacctggcct ggtctggcca ctctgccaga gtgggtgctg ccagggacat ggccagggct
     2761 ggtgtgtcca tccctgaaat catgcaggct ggtggctgga ccaatgtgaa catagtgatg
     2821 aactacatca gaaacctgga ctctgagact ggggccatgg tgaggctgct cgaggatggg
     2881 gactgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2941 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3001 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3061 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3121 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3181 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3241 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3301 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
     3361 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3421 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3481 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
     3541 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     3601 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     3661 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     3721 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     3781 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     3841 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     3901 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     3961 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     4021 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4081 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4141 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4201 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4261 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4441 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4561 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     4621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4681 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     4741 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4801 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4861 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4921 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5101 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5161 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5221 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5281 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5341 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5401 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5461 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5521 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5581 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5641 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5701 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5761 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5821 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5881 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6241 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6781 atgt
//
