LOCUS       sequence-317853-        7072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of  Kir2.1-2A-GFP in a Flp-dependent fashion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7072)
  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 7072)
  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-FRT Kir2.1-2A-GFP
FEATURES             Location/Qualifiers
     source          1..7072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..48
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      54..63
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             63..92
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1383..1436
                     /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      1446..1455
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1452..1454,1455..1457,1458..2171)
                     /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(1497..1518)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1758..1777)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2105..2126
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2287..2334)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(2412..2859)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   2904..3033
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3095..3683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3175..3194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3854..4645,4646..4714))
                     /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     4477..4496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4715..4819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4887..4905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4943..4965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5065..5084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5101..5556)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5238..5259)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5450..5469
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5631..5771
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5642..5771
                     /label=AAV2 ITR
     polyA_signal    5811..6287
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6069..6088
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6319..6907
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6425..6436
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6834..6854
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcgg cgcgccacca
       61 tggagcagaa gctgatcagc gaggaggacc tgggcagtgt gagaaccaac cggtacagca
      121 ttgtctcttc ggaggaagat ggcatgaagc tggccactat ggcagttgcc aatggctttg
      181 ggaatggcaa gagtaaagtc catacccgac aacagtgcag gagccgcttt gtgaagaaag
      241 atggtcattg caatgttcag tttatcaacg tgggtgagaa gggacagagg tacctggcag
      301 acatctttac tacctgtgtc gacatccgct ggaggtggat gctggttatc ttctgtcttg
      361 ccttcgtgct ctcctggctg ttctttggct gtgtgttttg gttgatagcc ctgctccatg
      421 gggatctaga tacttctaaa gtgagcaaag catgcgtgtc ggaggttaac agcttcacgg
      481 ctgccttcct cttctccatc gagacccaga caaccattgg ctatggtttc aggtgtgtga
      541 cagacgaatg cccaattgct gtcttcatgg tggtattcca gtcaattgta ggctgcatca
      601 ttgacgcctt catcattggt gcagtcatgg cgaagatggc aaagccaaag aagagaaatg
      661 agactcttgt cttcagtcac aatgctgtga ttgccatgag ggatggcaaa ctctgcttga
      721 tgtggagagt gggtaacctt cgaaagagcc accttgtggg agctcatgtc cgggcacagc
      781 ttctcaaatc taggatcact tcagaagggg agtttatccc tttggaccag atagacatca
      841 atgttggttt tgatagtgga attgaccgca tatttctagt gtcccccatc actatcgttc
      901 acgaaataga tgaagacagc cctttatatg acttgagtaa gcaggacatt gacaatgcag
      961 actttgaaat tgttgtcata ctggaaggca tggtggaggc gactgccatg acaactcaat
     1021 gccggagttc gtatctggcc aatgaaattc tctggggtca ccgctatgag ccagtgctct
     1081 ttgaagagaa acactactat aaagtagact attcaagatt ccataagact tatgaagtac
     1141 ctaacacccc cctttgtagt gccagagact tagcagagaa gaaatacatc ctttcaaatg
     1201 caaattcatt ttgctatgaa aatgaagttg ccctaacaag caaagaggaa gaggaggata
     1261 gtgagaacgg agtcccagag agcacaagca cagactcacc tcctggcata gatctccaca
     1321 accaggcaag cgtacctcta gagcccaggc ccttaaggcg agaatcggag ataggcagtg
     1381 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccagcgg
     1441 atcccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1501 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1561 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1621 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1681 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1741 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1801 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1861 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1921 agcagaagaa cggcatcaag gcgaacttca agatccgcca caacatcgag gacggcggcg
     1981 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2041 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2101 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2161 tttacaagta actcgacgaa gttcctatac cttttgaaga ataggaactt cggaatagga
     2221 acttctgtcg atggtgatgc ttggcaattc gggtcgatgg tgaagcattg gtcttcctat
     2281 tttgatgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcggtacc
     2341 cccgggcccg cggtatcaaa tcaattcttt gccaaaatga tgagacagca caggatccgg
     2401 taccttctcg actgcgctct caggcacgac acgactcctc cgctgcccac cgcagactga
     2461 ggcagcgctg agtcgccggc gccgcagcgc agatggtcgc gcccgtgccc ccctatctcg
     2521 cgcctcgcgt ggtgcggtcc ggctgggccg gcggcggcgc ggacgcgacc aaggtggccg
     2581 ggaaggggag tttgcggggg accggcgagt gacgtcagcg cgccttcagt gctgaggcgg
     2641 cggtggcgcg cgccgccagg cgggggcgaa ggcactgtcc gcggtgctga agctggcagt
     2701 gcgcacgcgc ctcgccgcat cctgtttccc ctccccctct ctgatagggg atgcgcaatt
     2761 tggggaatgg gggttgggtg cttgtccagt gggtcggggt cggtcgtcag gtaggcaccc
     2821 ccaccccgcc tcatcctggt cctaaaaccc acttgcactc atacgcaggg ccctctgcag
     2881 tctagacaca cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     2941 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct
     3001 cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg
     3061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     4921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5101 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     5161 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     5221 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5281 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     5341 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5401 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5461 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5521 cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg
     5581 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca
     5641 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     5701 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     5761 taggggttcc tgcggccgct cggtccgcac gtggttacct acaaaatcag aaggacaggg
     5821 aagggagcag tggttcacgc ctgtaatccc agcaatttgg gaggccaagg tgggtagatc
     5881 acctgagatt aggagttgga gaccagcctg gccaatatgg tgaaaccccg tctctaccaa
     5941 aaaaacaaaa attagctgag cctggtcatg catgcctgga atcccaacaa ctcgggaggc
     6001 tgaggcagga gaatcgcttg aacccaggag gcggagattg cagtgagcca agattgtgcc
     6061 actgcactcc agcttggttc ccaatagacc ccgcaggccc tacaggttgt cttcccaact
     6121 tgccccttgc tccataccac ccccctccac cccataatat tatagaagga cacctagtca
     6181 gacaaaatga tgcaacttaa ttttattagg acaaggctgg tgggcactgg agtggcaact
     6241 tccagggcca ggagaggcac tggggagggg tcacagggat gccacccgta gatctctcga
     6301 gcagcgctcg gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc
     6361 gaaggcgaag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg
     6421 ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa
     6481 cataggcgag cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag
     6541 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     6601 tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc
     6661 cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa
     6721 cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga
     6781 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
     6841 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     6901 gttgattatc gataagcttg atatcgaatt catcgatcgg ccgcatataa gcttgaagtt
     6961 cctattccga agttcctatt cttcaaaagg tataggaact tcgggtcgat ggtgagatct
     7021 ggactagagg gtcgatggtg atgcttggat caaaatagga agaatcccaa tg
//
