LOCUS       Exported                9325 bp ds-DNA     circular SYN 15-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9325)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 16, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9325
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2551..2556
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /label=source:recombinant plasmid
                     /note="/locus_tag=source:recombinant plasmid"
     misc_feature    1..413
                     /locus_tag="5' LTR"
                     /label=5' LTR
     misc_feature    464..601
                     /locus_tag="psi"
                     /label=psi
     misc_feature    568..932
                     /locus_tag="gag"
                     /label=gag
     misc_feature    1078..1319
                     /locus_tag="RRE"
                     /label=RRE
     misc_feature    1752..1869
                     /locus_tag="cPPT"
                     /label=cPPT
     promoter        1922..2297
                     /locus_tag="TRE3G"
                     /label=TRE3G
     misc_feature    2276..2297
                     /locus_tag="PCR primer"
                     /label=PCR primer
                     /note="Geneious type: PCR_primer"
     misc_feature    2302..2307
                     /locus_tag="NheI"
                     /label=NheI
     misc_feature    2308..2313
                     /locus_tag="Kozak sequence"
                     /label=Kozak sequence
     misc_feature    2314..2403
                     /locus_tag="AKAP1 transmembrane domain"
                     /label=AKAP1 transmembrane domain
     misc_feature    2404..2472
                     /locus_tag="mito linker"
                     /label=mito linker
     misc_feature    2473..2502
                     /locus_tag="GSlinker"
                     /label=GSlinker
     misc_feature    2503..2508
                     /locus_tag="BamHI"
                     /label=BamHI
     misc_feature    2509..2550
                     /locus_tag="V5 tag"
                     /label=V5 tag
     misc_feature    2551..2556
                     /locus_tag="KpnI"
                     /label=KpnI
     misc_feature    2557..3918
                     /locus_tag="LOV-Turbo, human codon optimized"
                     /label=LOV-Turbo, human codon optimized
     misc_feature    2707..2709
                     /locus_tag="T52S"
                     /label=T52S
     misc_feature    2770..2772
                     /locus_tag="L73Q"
                     /label=L73Q
     misc_feature    2782..3198
                     /locus_tag="LOV"
                     /label=LOV
     misc_feature    3394..3396
                     /locus_tag="I147T"
                     /label=I147T
     misc_feature    3451..3453
                     /locus_tag="A166V"
                     /label=A166V
     misc_feature    3646..3648
                     /locus_tag="I231V"
                     /label=I231V
     misc_feature    3811..3813
                     /locus_tag="F286L"
                     /label=F286L
     misc_feature    3919..3921
                     /locus_tag="Stop(1)"
                     /label=Stop
                     /label=Stop(1)
     misc_feature    3922..3924
                     /locus_tag="Stop"
                     /label=Stop
     misc_feature    3930..3935
                     /locus_tag="EcoRI"
                     /label=EcoRI
     misc_feature    3936..4524
                     /locus_tag="WPRE"
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(3989..4009)
                     /locus_tag="WPRE-R"
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4407..4418)
                     /codon_start=1
                     /locus_tag="Factor Xa site"
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4544..5054
                     /locus_tag="hPGK promoter"
                     /label=hPGK promoter
     CDS             5064..5807
                     /locus_tag="Tet-On 3G"
                     /label=Tet-On 3G
     misc_feature    5808..5873
                     /locus_tag="P2A"
                     /label=P2A
     misc_feature    5874..6467
                     /locus_tag="puro"
                     /label=puro
     misc_feature    6090..6096
                     /locus_tag="RR47"
                     /label=RR47
     misc_feature    6091..6096
                     /locus_tag="RR48"
                     /label=RR48
     misc_feature    6568..6803
                     /locus_tag="3' LTR"
                     /label=3' LTR
     misc_feature    6874..7004
                     /locus_tag="PolyA signal"
                     /label=PolyA signal
     misc_feature    7242..8102
                     /locus_tag="AmpR"
                     /label=AmpR
     misc_feature    8363..8978
                     /locus_tag="pUC/Origin"
                     /label=pUC/Origin
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga caattctcga cctcgagaca aatggcagta
     1741 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     1801 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     1861 caaaattttc gggtttatta cagggacagc agagatccac tttggccgcg aatcgatatg
     1921 ttttactccc tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag
     1981 aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta
     2041 tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca
     2101 gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag
     2161 aacgtataag ctttaggcgt gtacggtggg cgcctataaa agcagagctc gtttagtgaa
     2221 ccgtcagatc gcctggagca attccacaac acttttgtct tataccaact ttccgtacca
     2281 cttcctaccc tcgtaaatag agctagcgcc accatggcca tccagctgcg cagcctgttc
     2341 cccctggccc tgcccggcct gctggccctg ctgggctggt ggtggttctt cagccgcaag
     2401 aaggacctgg agctgaagct gcgcatcctg cagagcaccg tgccccgcgc ccgcgaccct
     2461 cccgtggcca ccggttccag tggaggcggt agctcaggtt cgggatccgg caagcccatc
     2521 cccaaccccc tgctgggcct ggacagcacc ggtaccaaag acaacacagt gcctctgaag
     2581 ctgatcgccc tcctggccaa tggagagttc cacagcggcg agcagctggg cgaaaccctg
     2641 ggcatgagca gagccgctat caacaagcac atccagaccc ttcgcgactg gggcgtcgac
     2701 gtgttcagcg tgcccggcaa gggctacagc ctgcctgagc ctatccccct gctgaacgcc
     2761 aagcagattc agggccagct ggaatttaga gccaccaccc tggaaagaat cgagaagtct
     2821 tttgtgatca ccgaccccag actgcctgat aatcctatca tcttcgtgtc tgatagcttc
     2881 ctgcagctca ccgagtattc tcgggaagag atcctcggaa gaaattgcag attcctgcag
     2941 ggcccagaga cagatagagc cacagtgcgg aagatcagag atgccatcga caaccagaca
     3001 gaggtgaccg tgcagctgat caactacacc aagtccggca aaaagttctg gaacgttttc
     3061 catctccaac ctatgagaga ctacaagggc gacgtgcagt acttcatcgg agttcagctg
     3121 gacggcaccg agagactgca cggcgccgca gagcgggaag ctgtgtgcct ggtcaagaag
     3181 accgccttcc agatcgccgt ggctgtgctg cccgtggtgg acagcacaaa ccagtacctg
     3241 ctggacagaa tcggcgagct gaagtccggc gacgcttgta tcgccgaata ccagcaggcc
     3301 ggcagaggca gcagaggccg gaaatggttc agccccttcg gcgccaacct gtacctgagc
     3361 atgttttggc ggctgaaaag aggccctgcc gccacaggcc tgggccccgt gataggaatc
     3421 gtgatggccg aggctctgag gaagctgggt gtggacaaag tgcgggtgaa gtggcctaac
     3481 gacctgtacc tgcaggacag aaagctggcc ggcatcctgg tggaactggc tggcatcacc
     3541 ggcgacgccg ctcagattgt gatcggcgct ggcatcaatg tggccatgag acgcgtggaa
     3601 gagagcgtgg tcaaccaggg ctggatcaca ctgcaagagg ccggtgtgaa cctggatagg
     3661 aacaccctgg ccgccaccct gatcagagag ctgagagccg ctctggaatt gtttgagcag
     3721 gagggcctgg ccccttacct gccacggtgg gagaaactgg ataacttcat caacaggcct
     3781 gtgaagctga tcatcggcga taaggaaatc ctgggaatca gccggggcat cgacaagcag
     3841 ggagccctgc tgctggaaca agatggagtg attaagcctt ggatgggcgg agaaatcagc
     3901 ctgcggtccg ccgagaagta ataggggccg aattcaatca acctctggat tacaaaattt
     3961 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4021 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4081 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4141 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4201 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4261 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4321 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     4381 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4441 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4501 tctccctttg ggccgcctcc ccgcctgtct gcagatatca tccggggttg gggttgcgcc
     4561 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     4621 aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc
     4681 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     4741 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     4801 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     4861 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
     4921 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
     4981 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
     5041 acctctctcc ccagcaattc accatgtcta gactggacaa gagcaaagtc ataaactctg
     5101 ctctggaatt actcaatgga gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa
     5161 agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg
     5221 atgccctgcc aatcgagatg ctggacaggc atcataccca ctcctgcccc ctggaaggcg
     5281 agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct ctcctctcac
     5341 atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc
     5401 tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg
     5461 ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag gagcatcaag
     5521 tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt ctgaaacaag
     5581 caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc ctggaactaa
     5641 tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg
     5701 acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc
     5761 tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccggggga agcggagcta
     5821 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga cctaccgagt
     5881 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg
     5941 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg
     6001 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     6061 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     6121 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     6181 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     6241 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     6301 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     6361 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     6421 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctaa ctgtttcgcc
     6481 tcggcgtccg gactagaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     6541 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     6601 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6661 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6721 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6781 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6841 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6961 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     7021 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     7081 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     7141 agaagtagtg aggaggcttt tttggaggcc tttcaaatat gtatccgctc atgagacaat
     7201 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7261 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7321 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7381 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7441 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7501 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7561 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     7621 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     7681 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     7741 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     7801 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     7861 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7921 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7981 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8041 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8101 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8161 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8221 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     8281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8401 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     8461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     8521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     8641 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     8701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     9001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     9061 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     9121 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     9181 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     9241 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     9301 gggaacaaaa gctggagctg caagc
//
