LOCUS       sequence-173363-        5509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination transfer plasmid for making AAV. Contains a DIO 
            cassette for cre-induction. For introducing large constructs into 
            AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5509)
  AUTHORS   Cammack AJ, Moudgil A, Chen J, Vasek MJ, Shabsovich M, McCullough K,
            Yen A, Lagunas T, Maloney SE, He J, Chen X, Hooda M, Wilkinson MN, 
            Miller TM, Mitra RD, Dougherty JD
  TITLE     A viral toolkit for recording transcription factor-DNA interactions 
            in live mouse tissues.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 5;117(18):10003-10014. doi: 
            10.1073/pnas.1918241117. Epub 2020 Apr 16.
  PUBMED    32300008
REFERENCE   2  (bases 1 to 5509)
  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"; doi: 
            "10.1073/pnas.1918241117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-05-5- 5"; volume: "117"; issue: "18"; pages: 
            "10003-10014"
COMMENT     Sequence Label: pAAV-short Cre-on G/W
FEATURES             Location/Qualifiers
     source          1..5509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(148..181)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(232..265)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    278..402
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(443..748)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(545..564)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1090..1749)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1664..1683
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1803..1809,1810..1827,1828..1833))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1858..1982)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1998..2031
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2082..2115
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2164..2181)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2170..2377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2415..2544
                     /label=AAV2 ITR
     terminator      2726..2769
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2826..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2906..2925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3585..4376,4377..4445))
                     /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     4208..4227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4446..4509)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4894..4916
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4908..4925
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4909..4925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4936..4955
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4936..4954
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   4991..5120
                     /label=AAV2 ITR
ORIGIN
        1 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
       61 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      121 gcgggcgggg agtcgctgca aacctctata acttcgtata ggatacttta tacgaagtta
      181 tgcagaatgg tagctggatt gtagctgcta ttagcaatat gaaacctctt aataacttcg
      241 tatagcatac attatacgaa gttatggcaa ctaaagtacc actttgtaca agaaagctga
      301 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
      361 gactacataa tactgtaaaa cacaacatat ccagtcacta tggtcgacct gcagactggc
      421 tgtgtataac ggagcctgac atttatattc cccagaacat caggttaatg gcgtttttga
      481 tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag accggcacac
      541 tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc accgggtaaa
      601 gttcacggga gactttatct gacagcagac gtgcactggc cagggggatc accatccgtc
      661 gcccgggcgt gtcaataata tcactctgta catccacaaa cagacgataa cggctctctc
      721 ttttataggt gtaaacctta aactgcattt caccagtccc tgttctcgtc agcaaaagag
      781 ccgttcattt caataaaccg ggcgacctca gccatccctt cctgattttc cgctttccag
      841 cgttcggcac gcagacgacg ggcttcattc tgcatggttg tgcttaccag accggagata
      901 ttgacatcat atatgccttg agcaactgat agctgtcgct gtcaactgtc actgtaatac
      961 gctgcttcat agcacacctc tttttgacat acttcgggta tacatatcag tatatattct
     1021 tataccgcaa aaatcagcgc gcaaatacgc atactgttat ctggctttta gtaagccgga
     1081 tccacgcgtt tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     1141 tctgccgaca tggaagccat cacagacggc atgatgaacc tgaatcgcca gcggcatcag
     1201 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     1261 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     1321 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     1381 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     1441 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     1501 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     1561 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     1621 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     1681 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     1741 tttctccatt ttagcttcct tagctcctga aaatctcgcc ggatcctaac tcaaaatcca
     1801 cacattatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg cggccgccat
     1861 agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa
     1921 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttttt gtacaaactt
     1981 gtctagcata agtgcagata acttcgtata aagtatccta tacgaagtta tttgccttaa
     2041 cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatgtat
     2101 gctatacgaa gttattttat ctccagagca tttaaatcac tggtctacta aaccgctgat
     2161 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2221 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2281 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2341 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atagagagtc
     2401 gatcgtgtga tcgaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2461 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2521 ctcagtgagc gagcgagcgc gcagcatgta gagactgggc ctcatgggcc ttccgcctat
     2581 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct
     2641 gtttccttgc gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg
     2701 ttcgggtaaa gcctggggtg cctaaataaa acgaaaggcc cagtcttccg actgagcctt
     2761 tcgttttatt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3241 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3721 ccgcgagaac cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4501 ggatacatat ttgaagctaa taacaggcct gctggtaatc gcaggccttt ttatttggtg
     4561 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4621 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4681 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4741 gggttgagtg gccgctacag ggcgctccca ttcgccattc aggctgcgca actgttggga
     4801 agggcgtttc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     4861 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     4921 ccagtgagcg cgacgtaata cgactcacta tagggcgaat tgtaggcgga aggccgtcaa
     4981 ggccgcatgg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     5041 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     5101 ctccatcact aggggttcct attgacgtca gatcgctaaa taatgacgta tgttcccata
     5161 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5221 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5281 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5341 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     5401 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     5461 ttattttgtg cagcgatggg ggcggggggg ggggkkrcsc gcgccaggc
//
