LOCUS       sequence-90593-e        6932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses diphtheria toxin A subunit in Cre positive cells, and 
            mCherry in all the infected cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6932)
  AUTHORS   Wu Z, Autry AE, Bergan JF, Watabe-Uchida M, Dulac CG
  TITLE     Galanin neurons in the medial preoptic area govern parental 
            behaviour.
  JOURNAL   Nature. 2014 May 15;509(7500):325-30. doi: 10.1038/nature13307.
  PUBMED    24828191
REFERENCE   2  (bases 1 to 6932)
  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"; journalName: 
            "Nature"; date: "2014-05-15- 15"; volume: "509"; issue: "7500"; 
            pages: "325-30"
COMMENT     Sequence Label: pAAV-mCherry-flex-dtA
FEATURES             Location/Qualifiers
     source          1..6932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1483..1492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1498..2205
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1640..1658)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1908..1927
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(2319..2891)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     misc_feature    3059..3647
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3112..3132)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3530..3541)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3679..4155
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3878..3897)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4195..4335
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4195..4324
                     /label=AAV2 ITR
     rep_origin      4410..4865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4497..4516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4707..4728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4882..4901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5001..5023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5061..5079)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5147..5251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5252..5320,5321..6112)
                     /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(5470..5489)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6283..6871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6772..6791
                     /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 cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 aattcattac ctcgtatagg atactttata cgaagttatg ccgccaccat gggtggggtg
     1501 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1561 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1621 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1681 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1741 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1801 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1861 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1921 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1981 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     2041 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     2101 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     2161 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaataacc tcgtatagca
     2221 tacattatac gaagtaatag atcttagagc tttaaatctc tgtaggtagt ttgtccaatt
     2281 atgtcacacc acagaagtaa ggttccttca caaagagatc gcctgacacg atttcctgca
     2341 caggcttgag ccatatactc atacatcgca tcttggccac gttttccacg ggtttcaaaa
     2401 ttaatctcaa gttctacgct taacgctttc gcctgttccc agttattaat atattcaacg
     2461 ctagaactcc cctcagcgaa gggaaggctg agcactacac gcgaagcacc atcaccgaac
     2521 cttttgataa actcttccgt tccgacttgc tccatcaacg gttcagtgag acttaaacct
     2581 aactctttct taatagtttc ggcattatcc acttttagtg cgagaacctt cgtcagtcct
     2641 ggatacgtca ctttgaccac gcctccagct tttccagaga gcgggttttc attatctaca
     2701 gagtatcccg cagcgtcgta tttattgtcg gtactataaa accctttcca atcatcgtca
     2761 taatttcctt gtgtaccaga ttttggcttt tgtatacctt tttgaatgga atctacataa
     2821 ccaggtttag tcccgtggta cgaagaaaag ttttccatca caaaagattt agaagaatca
     2881 acaacatcat caggatccat ggcgaggacg tcgacataac ttcgtataaa gtatcctata
     2941 cgaggtaatt tgccttaacc cagaaattat cagtactatt ctttagaatg gtgcaaagaa
     3001 ttacttcgta taatgtatgc tatacgaggt tatcgcatgc tagatcaagc ttatcgataa
     3061 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3121 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3181 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3241 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3301 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3361 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3421 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3481 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3541 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3601 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     3661 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     3721 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     3781 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     3841 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     3901 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     3961 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     4021 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     4081 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     4141 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     4201 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4261 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4321 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4381 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4441 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4501 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4561 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4621 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4681 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4741 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4801 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4861 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4921 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4981 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5041 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5101 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5161 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5221 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5281 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5341 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5401 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5461 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5521 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5581 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5641 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5701 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5761 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5821 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5881 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5941 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6001 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6061 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6121 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6181 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6241 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6301 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6361 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6421 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6481 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6541 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6601 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6661 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6721 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6781 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6841 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6901 cctggccttt tgctggcctt ttgctcacat gt
//
