LOCUS       sequence-317857-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Kir2.1 in cells not expressing Cre..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  AUTHORS   Beier KT, Kim CK, Hoerbelt P, Hung LW, Heifets BD, DeLoach KE, Mosca
            TJ, Neuner S, Deisseroth K, Luo L, Malenka RC
  TITLE     Rabies screen reveals GPe control of cocaine-triggered plasticity.
  JOURNAL   Nature. 2017 Sep 21;549(7672):345-350. doi: 10.1038/nature23888. 
            Epub 2017 Sep 13.
  PUBMED    28902833
REFERENCE   2  (bases 1 to 7016)
  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: 
            "10.1038/nature23888"; journalName: "Nature"; date: "2017-09-21- 
            21"; volume: "549"; issue: "7672"; pages: "345-350"
COMMENT     Sequence Label: pAAV hSyn FLEx-OFF Kir2.1-2A-GFP
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..34
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             47..76
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1367..1420
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1430..1439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1436..1438,1439..1441,1442..2155)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1481..1502)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1742..1761)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2089..2110
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2165..2198)
                     /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(2269..2302)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2350..2938
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2403..2423)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2821..2832)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2970..3446
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3169..3188)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3486..3626
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3486..3615
                     /label=AAV2 ITR
     rep_origin      3701..4156
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3788..3807)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3998..4019
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4173..4192)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4292..4314
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4352..4370)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4438..4542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4543..4611,4612..5403)
                     /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(4761..4780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5574..6162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6063..6082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6224..6353
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        6398..6845
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    6921..6954
                     /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)."
ORIGIN
        1 ataacttcgt ataatgtatg ctatacgaag ttatgtcgag accatggagc agaagctgat
       61 cagcgaggag gacctgggca gtgtgagaac caaccggtac agcattgtct cttcggagga
      121 agatggcatg aagctggcca ctatggcagt tgccaatggc tttgggaatg gcaagagtaa
      181 agtccatacc cgacaacagt gcaggagccg ctttgtgaag aaagatggtc attgcaatgt
      241 tcagtttatc aacgtgggtg agaagggaca gaggtacctg gcagacatct ttactacctg
      301 tgtcgacatc cgctggaggt ggatgctggt tatcttctgt cttgccttcg tgctctcctg
      361 gctgttcttt ggctgtgtgt tttggttgat agccctgctc catggggatc tagatacttc
      421 taaagtgagc aaagcatgcg tgtcggaggt taacagcttc acggctgcct tcctcttctc
      481 catcgagacc cagacaacca ttggctatgg tttcaggtgt gtgacagacg aatgcccaat
      541 tgctgtcttc atggtggtat tccagtcaat tgtaggctgc atcattgacg ccttcatcat
      601 tggtgcagtc atggcgaaga tggcaaagcc aaagaagaga aatgagactc ttgtcttcag
      661 tcacaatgct gtgattgcca tgagggatgg caaactctgc ttgatgtgga gagtgggtaa
      721 ccttcgaaag agccaccttg tgggagctca tgtccgggca cagcttctca aatctaggat
      781 cacttcagaa ggggagttta tccctttgga ccagatagac atcaatgttg gttttgatag
      841 tggaattgac cgcatatttc tagtgtcccc catcactatc gttcacgaaa tagatgaaga
      901 cagcccttta tatgacttga gtaagcagga cattgacaat gcagactttg aaattgttgt
      961 catactggaa ggcatggtgg aggcgactgc catgacaact caatgccgga gttcgtatct
     1021 ggccaatgaa attctctggg gtcaccgcta tgagccagtg ctctttgaag agaaacacta
     1081 ctataaagta gactattcaa gattccataa gacttatgaa gtacctaaca cccccctttg
     1141 tagtgccaga gacttagcag agaagaaata catcctttca aatgcaaatt cattttgcta
     1201 tgaaaatgaa gttgccctaa caagcaaaga ggaagaggag gatagtgaga acggagtccc
     1261 agagagcaca agcacagact cacctcctgg catagatctc cacaaccagg caagcgtacc
     1321 tctagagccc aggcccttaa ggcgagaatc ggagataggc agtggagagg gcagaggaag
     1381 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca gcggatcccg ccaccatggt
     1441 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1501 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1561 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1621 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1681 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1741 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1801 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1861 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1921 caaggcgaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     1981 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2041 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2101 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctttaca agtaaggcgc
     2161 gcctataact tcgtatagga tactttatac gaagttatca ttgggattct tcctattttg
     2221 atccaagcat caccatcgac cctctagtcc agatctcacc atcgacccat aacttcgtat
     2281 agcatacatt atacgaagtt ataagcttat atgcggccga tcgatgaatt cgatatcaag
     2341 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2401 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2461 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2521 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2581 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2641 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2701 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2761 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2821 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2881 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     2941 cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     3001 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     3061 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     3121 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     3181 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     3241 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     3301 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     3361 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     3421 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     3481 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3541 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3601 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     3661 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3721 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     3781 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3841 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3901 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3961 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4021 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4081 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4141 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4201 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4261 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4321 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4381 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     6241 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     6301 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     6361 cacgcgtgtg tctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
     6421 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
     6481 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
     6541 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
     6601 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
     6661 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
     6721 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
     6781 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
     6841 cgcagtcgag aaggtaccgg atcctgtgct gtctcatcat tttggcaaag aattgatttg
     6901 ataccgcggg cccggccggg ataacttcgt ataaagtatc ctatacgaag ttatatcaaa
     6961 ataggaagac caatgcttca ccatcgaccc gaattgccaa gcatcaccat cgacac
//
