LOCUS       pAAV-EF1a-fDIO-s        7442 bp DNA     circular SYN 08-JAN-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7442)
  AUTHORS   Salk Institute
  TITLE     Direct Submission
  JOURNAL   Exported Mar 7, 2024 from SnapGene 7.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7442
                     /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"
                     /note="color: #ffe4c4"
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /note="color: #------"
     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)"
                     /note="color: #31849b; direction: RIGHT"
     CDS             complement(1777..2472)
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     misc_feature    2476..2535
                     /label=ts
                     /note="color: #99cc00"
     gene            complement(2542..3414)
                     /label=stGtACR2
                     /note="color: #ff9900; direction: LEFT"
     regulatory      3411..3420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     protein_bind    complement(3427..3460)
                     /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)"
                     /note="color: #31849b; direction: LEFT"
     misc_feature    3569..4157
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(4040..4051)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4039"
                     /translation="IEGR"
     polyA_signal    4189..4665
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   4705..4845
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      4920..5375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5657..5761
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5762..6622
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5762 .. 5830 / #ccffcc / signal sequence
                        2: 5831 .. 6622 / #ccffcc
                     Cleavage site after base 5830"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6793..7381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 gcgccttact taaaccgcgt agcctctgga aaatcggtgg cgcatgagat
     1621 aaaagagtca atagaactca tggacctcat gtcgatcacg ccctctgttc ttgcaggaag
     1681 aggtgccaca ggggagggca ttgaggacat ttccagctcc tcagggggtt tactctgggg
     1741 ggccatctct ttggtgttca gaataggctg ggactgttta cctccatcac cagcgccacc
     1801 agtagagtat ctggccacag ccacctcgtg ctgctcgacg tatgtctcat cgtcggcctc
     1861 cttgattctt tccagtctgt ggtccacgtt gtagacgccg ggcatcttga ggttcgtagc
     1921 gggtttcttg gatctgtatg ttgtctcaag gttgcagatc aggtgtcctc ctcccacgag
     1981 cttcagagcc atgtcacatg cgccttccag tcctccgtca gcagggtaca tcgtctcggt
     2041 ggaggcctcc cagccgagtg ttttcttctg catcacaggg ccgttggctg ggaagttcac
     2101 ccctctaacc ttgacgttgt agatgaggca gccgtcctgg aggctggtgt cctgggtagc
     2161 ggtcagcacg cccccgtctt cgtatgtggt gactctctcc catgtgaagc cctcagggaa
     2221 ggactgctta aagaagtcgg ggatgcccgg agggtgcttg atgaaggttc tgctgccgta
     2281 catgaagctg gtagccagga tgtcgaaggc gaaggggaga gggccgccct cgacgacctt
     2341 gattctcatg gtctgggtgc cctcgtaggg cttgccttcg ccctcggatg tgcacttgaa
     2401 gtggtggttg ttcacggtgc cctccatgta cagcttcatg ggcatgttct ccttaatcag
     2461 ctcgctcacc atcaccacgt tgatgtcgat ctggtccagg gggatgtact cgccctcgct
     2521 ggtgatcctg ctcttggcgg cccttccaaa cattctgata ttgacttcgt cgttaggagg
     2581 ggcattcttc tcgtgattca ggccaaaagg atcgtcggct tcgggtttct cctgccctgg
     2641 cagacatgcg gggtcccact tcccttccag cactccccag gcagtgctcc acaggatcac
     2701 tccatatgta tttttgcaca ggccgtcagc cagcaggtac agaatggcgc tcacgttctc
     2761 gtggatcacg cagactccgg tgggtgaaat cagccacacg attgggaaca tagtccaagc
     2821 aataaagaag atcaggcgca gcatcttgat cctcaccacc accttctgtg ccagaggtgt
     2881 cccgacagat tcaaagccat acagccccac ttgaaagatt gtgaagacca cactgtactc
     2941 gaacaccagg caggtggccc caaagaaaaa gaacagccac ttcatatact ctgcgggaga
     3001 aattgtagcg gtgattccca tgaccactct aatgatggaa gcggcctgtg caatgggatt
     3061 cagtgggata cttttgtact tcaccagagc catgttagaa atctgtccca gcatgattgg
     3121 acaggtgcac agccaggagg ccattcggct ccaaggaaag aatctgccgt ccctcatctg
     3181 cagggttccg ttgccagtac tggccagtgc ataagtgata gattcgacaa agggcaggta
     3241 cacggactcc caagggacct tgggagccct gctaatcatg aagattgtct ggcagcctga
     3301 gaccacggcc cacaccagct gcagcagtga gacgaaggtg gagtcgattc tggacatatt
     3361 gtagcactcg gtgtgagttg atgcccattc cccatagacg acctgtgatg ccatggtggc
     3421 gctagcgaag ttcctatact ttctagagaa taggaacttc ttgccttaac ccagaaatta
     3481 tcactgttat tctttagaat ggtgcaaaga gaagttccta taccttttga agaataggaa
     3541 cttcgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3601 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3661 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3721 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3781 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3841 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3901 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3961 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     4021 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttcctcccc gcggcctgct
     4081 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4141 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     4201 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     4261 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     4321 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     4381 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     4441 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     4501 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     4561 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     4621 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     4681 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4741 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4801 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4861 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4981 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5101 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     5161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5221 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     5281 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5341 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5401 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5461 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5521 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5581 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5641 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5701 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5761 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5821 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5881 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5941 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6001 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6061 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6121 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6181 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6241 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6301 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6361 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6421 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6481 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6541 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6601 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6661 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6721 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6901 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7441 gt
//
