LOCUS       sequence-501089-        4335 bp    DNA     circular SYN 19-MAR-2026
DEFINITION  Cre and Flp dependent DTA (diptheria toxin) expressing system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4335)
  AUTHORS   Moore JD, Bachmann LC, McElvain LE, Pfaff SL, Dulac C
  TITLE     A genetically-defined population of amygdalofugal neurons promotes 
            suckling and early postnatal growth.
  JOURNAL   bioRxiv [Preprint]. 2025 Oct 18:2025.10.18.683193. doi: 
            10.1101/2025.10.18.683193.
  PUBMED    41279177
REFERENCE   2  (bases 1 to 4335)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Oct 18:2025.10.18.683193. doi: 
            10.1101/2025.10.18.683193."
COMMENT     Sequence Label: scAAV-DDIO-DTA
FEATURES             Location/Qualifiers
     source          1..4335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1190..1401
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    complement(1505..1538)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1842..1851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1861..1894
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1900..1933
                     /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)"
     protein_bind    complement(2390..2423)
                     /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)"
     primer_bind     2441..2460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2710..2727)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2710..2726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2719..2741)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2935..2954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3054..3076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3114..3132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3200..3304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3305..3373,3374..4165)
                     /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(3523..3542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaat agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct cgagatctag
     1021 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     1081 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     1141 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctagcgctg ggcagagcgc
     1201 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
     1261 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     1321 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     1381 gggtttgccg ccagaacaca gctggtaagt ttagtctttt tgtcttttat ttcaggtccc
     1441 ggatccggtg gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc tttacttctc
     1501 ctagtaccgt tcgtatagca tacattatac gaagttatgc atattattca aattgtggaa
     1561 ttcaatactt accttcgtca gtcctggata cgtcactttg accacgcctc cagcttttcc
     1621 agagagcggg ttttcattat ctacagagta tcccgcagcg tcgtatttat tgtcggtact
     1681 ataaaaccct ttccaatcat cgtcataatt tccttgtgta ccagattttg gcttttgtat
     1741 gcctttttga atggaatcta cataaccagg tttagtcccg tggtacgaag aaaagttttc
     1801 catcacaaaa gatttagaag aatcaacaac atcatcagga tccatggtgg cggcgctagc
     1861 ataaattgct ataatgtatg ctatacgaag ttatcctagg aagttcatat tctctagaaa
     1921 gtataggaac ttcggtacct tagagcttta aatctctgta ggtagtttgt ccaattatgt
     1981 cacaccacag aagtaaggtt ccttcacaaa gagatcgcct gacacgattt cctgcacagg
     2041 cttgagccat atactcatac atcgcatctt ggccacgttt tccacgggtt tcaaaattaa
     2101 tctcaagttc tacgcttaac gctttcgcct gttcccagtt attaatatat tcaacgctag
     2161 aactcccctc agcgaaggga aggctgagca ctacacgcga agcaccatca ccgaatcttt
     2221 tgataaactc ttccgttccg acttgctcca tcaacggttc agtgagactt aaacctaact
     2281 ctttcttaat agtttcggca ttatccactt ttagtgcgag aacctggaaa aaaagggaaa
     2341 aaaaatcgat tgttagtatt ttcaggtccc ataacccgcc tggcgcgccg aagttcatat
     2401 actttctaga gaataggaac ttctgcattg ctttatttgt gaaatttgtg atgctattgc
     2461 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2521 tatgtttcag gttcaggggg aggtgtggga ggttttttaa actagtccac tccctctttg
     2581 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2641 cgggcggcct cagtgagcga gcgagcgcgc agagagggac agatccgggc ccgcatgcgt
     2701 cgacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2761 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2821 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     2881 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     2941 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3001 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3061 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3121 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3181 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3241 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3301 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3361 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3421 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3481 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3541 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3601 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3661 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3721 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3781 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3841 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3901 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3961 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4021 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4081 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4141 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4201 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4261 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4321 gatcaaagga tcttc
//
