LOCUS       Exported                7038 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Flp-Dependent EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-fDIO EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  TITLE     INTRSECT
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7038)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 2, 2022 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7038
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="/dnas_title=Exported"
     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"
     protein_bind    1450..1483
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    1463..1470
                     /label=TCTAGAAA
                     /note="TCTAGAAA"
     misc_feature    1534..1567
                     /label=FRT (F5)
                     /note="FRT (F5)"
     misc_feature    1547..1554
                     /label=TTCAAAAG
                     /note="TTCAAAAG"
     misc_feature    complement(1580..3010)
                     /label=tdTomato
                     /note="tdTomato"
     protein_bind    complement(3023..3056)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3036..3043
                     /label=TCTAGAAA
                     /note="TCTAGAAA"
     misc_feature    3107..3140
                     /label=FRT (F5 )
                     /note="FRT (F5 )"
     misc_feature    3120..3127
                     /label=TTCAAAAG
                     /note="TTCAAAAG"
     misc_feature    3165..3753
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3218..3238)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3636..3647)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3785..4261
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3984..4003)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4301..4441
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4301..4430
                     /label=AAV2 ITR
     rep_origin      4516..4971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4603..4622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4813..4834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4988..5007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5107..5129
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5167..5185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5253..5357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5358..6218
                     /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(5576..5595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6389..6977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6878..6897
                     /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 ctctagagtc
     1441 gactccggag aagttcctat tctctagaaa gtataggaac ttcgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttagaagttc ctattcttca aaaggtatag
     1561 gaacttcggc gcgccattct tacttgtaca gctcgtccat gccgtacagg aacaggtggt
     1621 ggcggccctc ggagcgctcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga
     1681 tgtccagctt ggtgtccacg tagtagtagc cgggcagttg cacgggcttc ttggccatgt
     1741 agatggtctt gaactccacc aggtagtggc cgccgtcctt cagcttcagg gcctggtgga
     1801 tctcgccctt cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc tcccagccca
     1861 tggtcttctt ctgcattacg gggccgtcgg gggggaagtt ggtgccgcgc atcttcacct
     1921 tgtagatcag cgtgccgtcc tgcagggagg agtcctgggt cacggtcacc agaccgccgt
     1981 cctcgaagtt catcacgcgc tcccacttga agccctcggg gaaggacagc ttcttgtaat
     2041 cggggatgtc ggcggggtgc ttcacgtacg ccttggagcc gtacatgaac tggggggaca
     2101 ggatgtccca ggcgaagggc agggggccgc ccttggtcac cttcagcttg gcggtctggg
     2161 tgccctcgta ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcatgg
     2221 agccctccat gcgcaccttg aagcgcatga actctttgat gacggccatg ttgttgtcct
     2281 cggaggaggc ggtgccggag ctgccgctgc cggtgctgcc ggtgccatgc cccaggaaca
     2341 ggtggtggcg gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg
     2401 aggtgatgtc cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg
     2461 ccatgtagat ggtcttgaac tccaccaggt agtggccgcc gtccttcagc ttcagggcct
     2521 ggtggatctc gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc
     2581 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct
     2641 tcaccttgta gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac
     2701 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct
     2761 tgtaatcggg gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg
     2821 gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg
     2881 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     2941 tcatggagcc ctccatgcgc accttgaagc gcatgaactc tttgatgacc tcctcgccct
     3001 tgctcaccat ggtggcgcta gcgaagttcc tatactttct agagaatagg aacttcttgc
     3061 cttaacccag aaattatcac tgttattctt tagaatggtg caaagagaag ttcctatacc
     3121 ttttgaagaa taggaacttc gaattcgata tcaagcttat cgataatcaa cctctggatt
     3181 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3241 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3301 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3361 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3421 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3481 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3541 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3601 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3661 ctccccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3721 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     3781 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     3841 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     3901 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     3961 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4021 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     4081 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4141 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4201 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4261 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4321 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4381 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4441 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4501 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4561 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4621 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4681 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4741 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4801 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4861 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4921 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4981 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5041 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5101 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5161 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5221 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5281 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5341 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5401 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5461 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5521 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5581 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5641 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5701 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5761 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5821 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5881 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5941 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6001 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6061 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6121 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6181 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6241 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6301 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6361 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6421 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6481 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6541 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6601 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6661 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6721 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6781 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6841 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6901 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6961 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7021 ggccttttgc tcacatgt
//
