LOCUS       sequence-374764-        6144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent FlpO expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  AUTHORS   Winnubst J, Bas E, Ferreira TA, Wu Z, Economo MN, Edson P, Arthur 
            BJ, Bruns C, Rokicki K, Schauder D, Olbris DJ, Murphy SD, Ackerman 
            DG, Arshadi C, Baldwin P, Blake R, Elsayed A, Hasan M, Ramirez D, 
            Dos Santos B, Weldon M, Zafar A, Dudman JT, Gerfen CR, Hantman AW, 
            Korff W, Sternson SM, Spruston N, Svoboda K, Chandrashekar J
  TITLE     Reconstruction of 1,000 Projection Neurons Reveals New Cell Types 
            and Organization of Long-Range Connectivity in the Mouse Brain.
  JOURNAL   Cell. 2019 Sep 19;179(1):268-281.e13. doi: 
            10.1016/j.cell.2019.07.042. Epub 2019 Sep 5.
  PUBMED    31495573
REFERENCE   2  (bases 1 to 6144)
  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: 
            "10.1016/j.cell.2019.07.042"; journalName: "Cell"; date: 
            "2019-09-19- 19"; volume: "179"; issue: "1"; pages: "268-281.e13"
COMMENT     Sequence Label: pAAV-Syn-Flex-FlpO
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   842..971
                     /label=AAV2 ITR
     intron          1564..1696
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1679..1698
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1709..1742
                     /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    1805..1838
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3185..3218)
                     /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(3289..3322)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3394..3982
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3447..3467)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3865..3876)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4060..4181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4071..4090)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4125..4144
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4343..4472
                     /label=AAV2 ITR
     rep_origin      4544..4999
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4631..4650)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4841..4862
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5025..5129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5130..5198,5199..5990)
                     /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(5348..5367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      841 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
      901 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      961 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
     1021 taggaagatc cgagctccag tgtgctggaa ttcgcccttt tcagtattta aattgacgac
     1081 cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc
     1141 agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc
     1201 gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg
     1261 cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt
     1321 ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg
     1381 gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag
     1441 cggaggagtc gtgtcgtgcc tgagagcgca gctgtgctcc tgggcaccgc gcagtccgcc
     1501 cccgcggctc ctggccagac cacccctagg accccctgcc ccaagtcgca gccgttggat
     1561 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     1621 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     1681 gcctttctct ccacaggtga tgatccccat aacttcgtat aaagtatcct atacgaagtt
     1741 atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc atcaccatcg
     1801 acacataact tcgtataatg tatgctatac gaagttatac tagtgctagc tgatcaacgc
     1861 gtcttaagga tatcaccggt tcagatccgc ctgttgatgt agctgctcag gtagtccagc
     1921 acctcctggc tgatgatgcc gttccaggcg gggtatctga tgctgccctc ggcgctgccc
     1981 ttcagctgct cgatgtgctg ccactcctcg atggggttgg tctcgtcctt cagggcgatc
     2041 atctccttgc tgatggggtc gtaggcgtag tacctggaca ccagggcgaa gtagtggtcg
     2101 gggatggcgg tgatctggtg ggtgtaggtg gtcctggcca cggcggaggc cctcttgtcg
     2161 ctccagttgc ccaccacgtt tgtcagctcg gtcaggccct tcatgctcag aaagctggtc
     2221 atcaggtgcc tgccgatgtg gctcttaggg ccgttcttga tagcgaagat ggggtagggg
     2281 gcgttcttct tcagggcctt gttgtagctg cgcaccaggt tgtccttcag cagctggtac
     2341 tcctgcttgt tgctgctgct gttgccggtc ctgttcactc tcttcagcac gggctcgctg
     2401 ttcctcagga actcgtccag gtacaccagg gggtcgatcc tgcctctggc gctgaaaaag
     2461 tagatgtgcc tggacacgct tgtcttggtc tcggtcacca ggcactgaat gatcacgccc
     2521 aggtacttgt tctgcaccag cttgaagctc ttggggtcca cgttcttgat gtcgctgaac
     2581 ctgccgcagt tgatgaatgt ggccaggaac aggaactggt acagggtctt ggtcttggtg
     2641 aacctgctgg tgtactcgaa gctgttcagg atcttctcgg tgatctccca gatgctctcg
     2701 ccctcggaca gcagggcctt cagcatcttc ttgctgtggc tgttgccctt gtcggcctcc
     2761 tcgctgctct cgaactgcag ctgcaggctg gacacgatgt cggtgatgtc gctctggtgc
     2821 ttctggccgt tgtaagggat gatggtgaac tcccaggcgg ggatcagctt cttcaggctg
     2881 gcctccagga tggtggcctt ctgggtcttg tacttgaact gcaggctctt gttcacgatg
     2941 tcgaagctca ggctgttgct gatgatggtg ttgtagctca tgaaggtggc cctcttgatg
     3001 gcggtgccgt tgtgggtgat catccagcac aggtaggtca gctcggcggc acagctggcg
     3061 atcttctcgc cgctgggcct ctcgaatctc tccacgaact gccgcaccag caccttgggg
     3121 ggggtcttgc acaggatgtc gaactggctc atggtggcgg atccggcgcg cccatatgac
     3181 tagtataact tcgtatagga tactttatac gaagttatca ttgggattct tcctattttg
     3241 atccaagcat caccatcgac cctctagtcc agatctcacc atcgacccat aacttcgtat
     3301 agcatacatt atacgaagtt atgtccctcg aagaggttcg aaacactgaa aagggcgaat
     3361 tctgcagatg atccactcga gaagcttatc gataatcaac ctctggatta caaaatttgt
     3421 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3481 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3541 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3601 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3661 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3721 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3781 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3841 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3901 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3961 tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct cgagcatgca
     4021 tctagagtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     4081 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     4141 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     4201 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     4261 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     4321 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     4381 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4441 cgggcggcct cagtgagcga gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc
     4501 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     4561 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4621 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4681 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4741 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4801 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4861 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4921 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4981 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5041 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5101 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5161 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5221 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5281 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5341 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5401 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5461 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5521 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5581 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5641 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5701 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5761 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5821 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5881 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5941 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6001 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6061 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6121 actgagcgtc agaccccgta gaaa
//
