LOCUS       sequence-222582-        6922 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 6922)
  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 6922)
  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..6922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        92..1270
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          323..1261
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1218..1238
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1334..1343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1349..2056
                     /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(1491..1509)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1759..1778
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(2170..2742)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     misc_feature    2910..3498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2963..2983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3381..3392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3530..4006
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3729..3748)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4046..4186
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4046..4175
                     /label=AAV2 ITR
     rep_origin      4261..4716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4348..4367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4558..4579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4733..4752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4852..4874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4912..4930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4998..5102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5103..5171,5172..5963)
                     /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(5321..5340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6134..6722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6623..6642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6784..6913
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
       61 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
      121 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      181 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      241 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      301 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      361 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
      421 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
      481 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
      541 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
      601 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
      661 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
      721 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
      781 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
      841 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
      901 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
      961 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     1021 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     1081 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     1141 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     1201 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     1261 ggtgtcgtga ggtaccggat cctctagagt caattcatta cctcgtatag gatactttat
     1321 acgaagttat gccgccacca tgggtggggt gagcaagggc gaggaggata acatggccat
     1381 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1441 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     1501 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     1561 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     1621 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     1681 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     1741 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     1801 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     1861 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     1921 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     1981 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     2041 cgagctgtac aagtaataac ctcgtatagc atacattata cgaagtaata gatcttagag
     2101 ctttaaatct ctgtaggtag tttgtccaat tatgtcacac cacagaagta aggttccttc
     2161 acaaagagat cgcctgacac gatttcctgc acaggcttga gccatatact catacatcgc
     2221 atcttggcca cgttttccac gggtttcaaa attaatctca agttctacgc ttaacgcttt
     2281 cgcctgttcc cagttattaa tatattcaac gctagaactc ccctcagcga agggaaggct
     2341 gagcactaca cgcgaagcac catcaccgaa ccttttgata aactcttccg ttccgacttg
     2401 ctccatcaac ggttcagtga gacttaaacc taactctttc ttaatagttt cggcattatc
     2461 cacttttagt gcgagaacct tcgtcagtcc tggatacgtc actttgacca cgcctccagc
     2521 ttttccagag agcgggtttt cattatctac agagtatccc gcagcgtcgt atttattgtc
     2581 ggtactataa aaccctttcc aatcatcgtc ataatttcct tgtgtaccag attttggctt
     2641 ttgtatacct ttttgaatgg aatctacata accaggttta gtcccgtggt acgaagaaaa
     2701 gttttccatc acaaaagatt tagaagaatc aacaacatca tcaggatcca tggcgaggac
     2761 gtcgacataa cttcgtataa agtatcctat acgaggtaat ttgccttaac ccagaaatta
     2821 tcagtactat tctttagaat ggtgcaaaga attacttcgt ataatgtatg ctatacgagg
     2881 ttatcgcatg ctagatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2941 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3001 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3061 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3121 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3181 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3241 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3301 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     3361 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3421 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3481 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     3541 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3601 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3661 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3721 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3781 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3841 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3901 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3961 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     4021 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4081 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4141 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4201 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4261 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4321 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4381 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4441 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4501 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4561 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4621 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4681 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4741 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4801 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4861 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4921 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4981 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5041 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5101 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5161 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5221 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5281 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5341 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5401 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5461 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5521 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5581 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5641 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5701 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5761 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5821 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5881 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5941 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6001 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6061 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6241 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6781 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     6841 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     6901 actaggggtt cctgcggccg ca
//
