LOCUS       Exported                5504 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  For AAV-TRE-Cre based Supernova, pK170 should be used with pK168 
            etc..
ACCESSION   .
VERSION     .
KEYWORDS        pK170.AAV-TRE-Cre-WPRE
            ova)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  AUTHORS   Luo W, Mizuno H, Iwata R, Nakazawa S, Yasuda K, Itohara S, Iwasato T
  TITLE     Supernova: A Versatile Vector System for Single-Cell Labeling and 
            Gene Function Studies in vivo.
  JOURNAL   Sci Rep. 2016 Oct 24;6:35747. doi: 10.1038/srep35747.
  PUBMED    27775045
REFERENCE   2  (bases 1 to 5504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        196..510
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    204..222
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    240..258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    275..293
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    311..329
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    347..365
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    382..400
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    418..436
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     483..507
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             544..564
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             568..1593
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS
                     VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR
                     PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG
                     IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW
                     ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY
                     LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG
                     D"
     misc_feature    1631..2219
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1684..1704)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2102..2113)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2251..2727
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2450..2469)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2767..2907
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2767..2896
                     /label=AAV2 ITR
     rep_origin      2982..3437
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3069..3088)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3279..3300
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3454..3473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3573..3595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3633..3651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3719..3823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3824..4684
                     /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(4042..4061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4855..5443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5344..5363
                     /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 cgcgtggcgc gccaggcgta tcacgaggcc
      181 ctttcgtctt cactcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact
      241 ccctatcagt gatagagaac gatgtcgagt ttactcccta tcagtgatag agaacgtatg
      301 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      361 agagaacgta tgtcgagttt atccctatca gtgatagaga acgtatgtcg agtttactcc
      421 ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc
      481 agagctcgtt tagtgaaccg tcagatcgcc tggagaattc tcgctacact gtcgaccatg
      541 ggcccaaaga agaagagaaa ggtttcgaat ttactgaccg tacaccaaaa tttgcctgca
      601 ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat
      661 cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg
      721 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
      781 tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc
      841 cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt
      901 tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag
      961 gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat
     1021 cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt
     1081 atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg
     1141 ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt
     1201 agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat
     1261 ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc
     1321 agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac
     1381 ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc
     1441 ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt
     1501 ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg
     1561 gcaatggtgc gcctgctgga agatggcgat tagggatatc agatccgcta gcgtcgacaa
     1621 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1681 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1741 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1801 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1861 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     1921 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1981 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2041 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2101 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2161 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2221 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     2281 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     2341 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     2401 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     2461 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     2521 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     2581 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     2641 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     2701 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     2761 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2821 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2881 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2941 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3001 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3061 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3121 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3181 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3241 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3301 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3361 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3421 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3481 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3541 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3601 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3661 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3721 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3781 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3841 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3901 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3961 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4021 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4081 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4141 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4201 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4261 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4321 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4381 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4441 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4501 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4561 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4621 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4681 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4741 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4801 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4861 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4921 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4981 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5041 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5101 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5161 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5221 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5281 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5341 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5401 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5461 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
