LOCUS       sequence-496015-        4398 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 4398)
  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 4398)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..4398
                     /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)."
     primer_bind     1583..1602
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1842..1863
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1905..1914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1924..1957
                     /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    1963..1996
                     /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     complement(2048..2069)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2453..2486)
                     /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     2504..2523
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2773..2790)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2773..2789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2782..2804)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2998..3017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3117..3139
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3177..3195)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3263..3367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3368..3436,3437..4228)
                     /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(3586..3605)
                     /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 cgcccgggca aagcccgggc 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 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     1621 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     1681 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     1741 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     1801 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     1861 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctccccatgg tggcggcgct
     1921 agcataaatt gctataatgt atgctatacg aagttatcct aggaagttca tattctctag
     1981 aaagtatagg aacttcggta cctcacttgt acagctcgtc catgccgaga gtgatcccgg
     2041 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     2101 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     2161 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     2221 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     2281 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     2341 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctgg aaaaaaaggg
     2401 aaaaaaaatc gattgttagt attttcaggt cccataaccc gcctggcgcg ccgaagttca
     2461 tatactttct agagaatagg aacttctgca ttgctttatt tgtgaaattt gtgatgctat
     2521 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2581 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaactagtc cactccctct
     2641 ttgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2701 gcccgggcgg cctcagtgag cgagcgagcg cgcagagagg gacagatccg ggcccgcatg
     2761 cgtcgacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2821 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2881 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     2941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     3001 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3061 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3121 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3181 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3241 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3301 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3361 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3421 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3481 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3541 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3601 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3661 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3721 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3781 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3841 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3901 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3961 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4021 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4081 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4141 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4201 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4261 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4321 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4381 aaagatcaaa ggatcttc
//
