LOCUS       sequence-177479-        6704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp recombinase dependent on GFP (Flp-DOG) for rAAV production and 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  AUTHORS   Tang JC, Drokhlyansky E, Etemad B, Rudolph S, Guo B, Wang S, Ellis 
            EG, Li JZ, Cepko CL
  TITLE     Detection and manipulation of live antigen-expressing cells using 
            conditionally stable nanobodies.
  JOURNAL   Elife. 2016 May 20;5. pii: e15312. doi: 10.7554/eLife.15312.
  PUBMED    27205882
REFERENCE   2  (bases 1 to 6704)
  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: "Elife."; 
            date: "2016-05-20"; volume: "5. pii: e15312. doi"; pages: " 
            10.7554/eLife.15312"
COMMENT     Sequence Label: pAAV-EF1a-Flp-DOG-NW
FEATURES             Location/Qualifiers
     source          1..6704
                     /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"
     polyA_signal    3451..3927
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3650..3669)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3967..4107
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3967..4096
                     /label=AAV2 ITR
     rep_origin      4182..4637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4269..4288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4479..4500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4654..4673)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4773..4795
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4833..4851)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4919..5023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5024..5092,5093..5884)
                     /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(5242..5261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6055..6643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6544..6563
                     /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 gccgacgtgc agctcgtgga atccggagga gcactggtcc agcctggggg atcactgaga
     1501 ctgtcttgtg tcgcttctgg ctttcctgtg aatcggtact ctatgagatg gtacaggcag
     1561 gcacctggaa aagagcgaga atgggtcgcc ggaatgtcat ctgctggcga taggagttcc
     1621 tacgtggata gcgtgaaagg acggttcacc attagaaggg acgatgctag aaacaccgtg
     1681 tacctccaga tgaactccct gaaacctgag gacactgctg tgtactacta caacgtcaat
     1741 gtgggctttg aatactgggg acatggcaca caagtgactg ttagttttgc atgcatggct
     1801 gacgtccaac ttgttgagag cggtggagca ctggtccagc ctgggggatc actgagactg
     1861 tcttgtgtcg cttctggctt tcctgtgaat cggtactcta tgagatggta caggcaggca
     1921 cctggaaaag agcgagaatg ggtcgccgga atgtcatctg ctggcgatag gagttcctac
     1981 gtggatagcg tgaaaggacg gttcaccatt agaagggacg atgctagaaa caccgtgtac
     2041 ctccagatga actccctgaa acctgaggac actgctgtgt actactacaa cgtcaatgtg
     2101 ggctttgaat actggggcca tgggacacag gttactgtta gcttcgcatg catgagccag
     2161 ttcgacatcc tgtgcaagac cccccccaag gtgctggtgc ggcagttcgt ggagagattc
     2221 gagaggccca gcggcgagaa gatcgccagc tgtgccgccg agctgaccta cctgtgctgg
     2281 atgatcaccc acaacggcac cgccatcaag agggccacct tcatgagcta caacaccatc
     2341 atcagcaaca gcctgagctt cgacatcgtg aacaagagcc tgcagttcaa gtacaagacc
     2401 cagaaggcca ccatcctgga ggccagcctg aagaagctga tccccgcctg ggagttcacc
     2461 atcatccctt acaacggcca gaagcaccag agcgacatca ccgacatcgt gtccagcctg
     2521 cagctgcagt tcgagagcag cgaggaggcc gacaagggca acagccacag caagaagatg
     2581 ctgaaggccc tgctgtccga gggcgagagc atctgggaga tcaccgagaa gatcctgaac
     2641 agcttcgagt acaccagcag gttcaccaag accaagaccc tgtaccagtt cctgttcctg
     2701 gccacattca tcaactgcgg caggttcagc gacatcaaga acgtggaccc caagagcttc
     2761 aagctggtgc agaacaagta cctgggcgtg atcattcagt gcctggtgac cgagaccaag
     2821 acaagcgtgt ccaggcacat ctactttttc agcgccagag gcaggatcga ccccctggtg
     2881 tacctggacg agttcctgag gaacagcgag cccgtgctga agagagtgaa caggaccggc
     2941 aacagcagca gcaacaagca ggagtaccag ctgctgaagg acaacctggt gcgcagctac
     3001 aacaaggccc tgaagaagaa cgccccctac cccatcttcg ctatcaagaa cggccctaag
     3061 agccacatcg gcaggcacct gatgaccagc tttctgagca tgaagggcct gaccgagctg
     3121 acaaacgtgg tgggcaactg gagcgacaag agggcctccg ccgtggccag gaccacctac
     3181 acccaccaga tcaccgccat ccccgaccac tacttcgccc tggtgtccag gtactacgcc
     3241 tacgacccca tcagcaagga gatgatcgcc ctgaaggacg agaccaaccc catcgaggag
     3301 tggcagcaca tcgagcagct gaagggcagc gccgagggca gcatcagata ccccgcctgg
     3361 aacggcatca tcagccagga ggtgctggac tacctgagca gctacatcaa caggcggatc
     3421 taggaattcg atgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3481 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3541 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3601 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3661 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3721 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3781 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3841 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3901 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3961 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4021 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4081 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4141 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4201 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4261 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4321 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4381 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4441 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4501 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4561 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4621 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4681 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4741 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4801 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4861 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5041 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5161 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5221 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5401 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5581 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5641 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5881 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5941 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6001 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6061 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6181 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
