LOCUS       sequence-317850-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of  Kir2.1-2A-GFP in a Cre-dependent fashion.
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-loxp Kir2.1-2A-GFP
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..30
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1321..1374
                     /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      1384..1393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1390..1392,1393..1395,1396..2109)
                     /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(1435..1456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1696..1715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2043..2064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2116..2149)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2212..2245)
                     /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)."
     promoter        complement(2321..2768)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   2813..2942
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3004..3592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3084..3103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3763..4554,4555..4623))
                     /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     4386..4405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4624..4728)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4796..4814
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4852..4874)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4974..4993
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5010..5465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5147..5168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5359..5378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5540..5680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5551..5680
                     /label=AAV2 ITR
     polyA_signal    5720..6196
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     5978..5997
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6228..6816
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6334..6345
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6743..6763
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    6864..6897
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6968..7001
                     /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)."
     regulatory      join(7008..7016,1)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gagcagaagc tgatcagcga ggaggacctg ggcagtgtga gaaccaaccg gtacagcatt
       61 gtctcttcgg aggaagatgg catgaagctg gccactatgg cagttgccaa tggctttggg
      121 aatggcaaga gtaaagtcca tacccgacaa cagtgcagga gccgctttgt gaagaaagat
      181 ggtcattgca atgttcagtt tatcaacgtg ggtgagaagg gacagaggta cctggcagac
      241 atctttacta cctgtgtcga catccgctgg aggtggatgc tggttatctt ctgtcttgcc
      301 ttcgtgctct cctggctgtt ctttggctgt gtgttttggt tgatagccct gctccatggg
      361 gatctagata cttctaaagt gagcaaagca tgcgtgtcgg aggttaacag cttcacggct
      421 gccttcctct tctccatcga gacccagaca accattggct atggtttcag gtgtgtgaca
      481 gacgaatgcc caattgctgt cttcatggtg gtattccagt caattgtagg ctgcatcatt
      541 gacgccttca tcattggtgc agtcatggcg aagatggcaa agccaaagaa gagaaatgag
      601 actcttgtct tcagtcacaa tgctgtgatt gccatgaggg atggcaaact ctgcttgatg
      661 tggagagtgg gtaaccttcg aaagagccac cttgtgggag ctcatgtccg ggcacagctt
      721 ctcaaatcta ggatcacttc agaaggggag tttatccctt tggaccagat agacatcaat
      781 gttggttttg atagtggaat tgaccgcata tttctagtgt cccccatcac tatcgttcac
      841 gaaatagatg aagacagccc tttatatgac ttgagtaagc aggacattga caatgcagac
      901 tttgaaattg ttgtcatact ggaaggcatg gtggaggcga ctgccatgac aactcaatgc
      961 cggagttcgt atctggccaa tgaaattctc tggggtcacc gctatgagcc agtgctcttt
     1021 gaagagaaac actactataa agtagactat tcaagattcc ataagactta tgaagtacct
     1081 aacacccccc tttgtagtgc cagagactta gcagagaaga aatacatcct ttcaaatgca
     1141 aattcatttt gctatgaaaa tgaagttgcc ctaacaagca aagaggaaga ggaggatagt
     1201 gagaacggag tcccagagag cacaagcaca gactcacctc ctggcataga tctccacaac
     1261 caggcaagcg tacctctaga gcccaggccc ttaaggcgag aatcggagat aggcagtgga
     1321 gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccagcggat
     1381 cccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1441 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1501 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1561 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1621 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1681 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1741 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1801 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1861 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
     1921 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1981 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2041 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2101 tacaagtaag tcgacataac ttcgtatagc atacattata cgaagttatg tgtcgatggt
     2161 gatgcttggc aattcgggtc gatggtgaag cattggtctt cctattttga tataacttcg
     2221 tataggatac tttatacgaa gttatcccgg ccgggcccgc ggtatcaaat caattctttg
     2281 ccaaaatgat gagacagcac aggatccggt accttctcga ctgcgctctc aggcacgaca
     2341 cgactcctcc gctgcccacc gcagactgag gcagcgctga gtcgccggcg ccgcagcgca
     2401 gatggtcgcg cccgtgcccc cctatctcgc gcctcgcgtg gtgcggtccg gctgggccgg
     2461 cggcggcgcg gacgcgacca aggtggccgg gaaggggagt ttgcggggga ccggcgagtg
     2521 acgtcagcgc gccttcagtg ctgaggcggc ggtggcgcgc gccgccaggc gggggcgaag
     2581 gcactgtccg cggtgctgaa gctggcagtg cgcacgcgcc tcgccgcatc ctgtttcccc
     2641 tccccctctc tgatagggga tgcgcaattt ggggaatggg ggttgggtgc ttgtccagtg
     2701 ggtcggggtc ggtcgtcagg taggcacccc caccccgcct catcctggtc ctaaaaccca
     2761 cttgcactca tacgcagggc cctctgcagt ctagacacac gcgtgcggcc gcaggaaccc
     2821 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2881 ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2941 ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3001 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3061 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3421 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3541 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3721 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3841 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3961 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4201 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4261 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4441 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4621 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4681 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4741 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4801 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4861 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4921 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     4981 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     5041 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     5101 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     5161 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5221 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     5281 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5341 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
     5401 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5461 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5521 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     5581 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     5641 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg
     5701 tggttaccta caaaatcaga aggacaggga agggagcagt ggttcacgcc tgtaatccca
     5761 gcaatttggg aggccaaggt gggtagatca cctgagatta ggagttggag accagcctgg
     5821 ccaatatggt gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc ctggtcatgc
     5881 atgcctggaa tcccaacaac tcgggaggct gaggcaggag aatcgcttga acccaggagg
     5941 cggagattgc agtgagccaa gattgtgcca ctgcactcca gcttggttcc caatagaccc
     6001 cgcaggccct acaggttgtc ttcccaactt gccccttgct ccataccacc cccctccacc
     6061 ccataatatt atagaaggac acctagtcag acaaaatgat gcaacttaat tttattagga
     6121 caaggctggt gggcactgga gtggcaactt ccagggccag gagaggcact ggggaggggt
     6181 cacagggatg ccacccgtag atctctcgag cagcgctcgg tatcgatgcg gggaggcggc
     6241 ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc
     6301 cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa
     6361 ggacgtcccg cgcagaatcc aggtggcaac ataggcgagc agccaaggaa aggacgatga
     6421 tttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg
     6481 ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc
     6541 ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac
     6601 agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa
     6661 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     6721 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     6781 caatctttca caaattttgt aatccagagg ttgattatcg ataagcttga tatcgaattc
     6841 atcgatcggc cgcatataag cttataactt cgtataatgt atgctatacg aagttatggg
     6901 tcgatggtga gatctggact agagggtcga tggtgatgct tggatcaaaa taggaagaat
     6961 cccaatgata acttcgtata aagtatccta tacgaagtta taggcgcgcc accatg
//
