LOCUS       Exported                7342 bp ds-DNA     circular SYN 08-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-DIO-Ace-mNeon2-Kv2.1PR
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7342)
  AUTHORS   Pieribone Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 9, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7342
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        134..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..792
                     /label=chicken beta-actin promoter
     intron          794..1811
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1678..1687
                     /label=1DIO-ANv3-Kv2.1
     protein_bind    1878..1911
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    1962..1995
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(1982..2004)
                     /label=2DIOAce-noKv
     primer_bind     1987..2023
                     /label=1DIO-ANv3-Kv2.1
     CDS             complement(2002..3636)
                     /codon_start=1
                     /label=Ace-mNeon2-Kv2.1PR
                     /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN
                     IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYISWVLTTPLLLLDLIVMTKMGGVMI
                     SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY
                     HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNEAILMGILDVLAKPLFGMGCLIAH
                     ETIFKSYPATHELHIFGSINGVDFDMVGQGTGNPNDGYEELNLKSTKGDLQFSPWILVP
                     HIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDGASLTVNYRYTYEGSHIKGE
                     AQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIISTFKWSYTTGNGKRYRSTART
                     TYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQKAFTDVMGMDELYKKSRITS
                     EGEYIPLDQIDINVFCYENEVQSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTE
                     GVIDMRSMSSIDSFISCATDFPEATRF"
     primer_bind     2002..2024
                     /label=2Kv2.1px
     primer_bind     complement(2176..2214)
                     /label=1Kv2.1px
     primer_bind     2200..2218
                     /label=1DIOAce-noKv
     primer_bind     2200..2218
                     /label=2AcTSER_inv
     primer_bind     2200..2215
                     /label=2AAV-Syn-TS/ER_ER1
     primer_bind     2281..2301
                     /label=280 - 2sfGR_AoIF2
     primer_bind     2281..2299
                     /label=2NLS-mCh
     primer_bind     complement(2921..2964)
                     /label=1_delPA
     primer_bind     complement(2921..2961)
                     /label=1del_A231
     primer_bind     complement(2921..2961)
                     /label=1A231_D
     primer_bind     complement(2921..2961)
                     /label=1A231_C
     primer_bind     complement(2921..2961)
                     /label=1A231_B
     primer_bind     complement(2921..2961)
                     /label=1A231_A
     primer_bind     complement(2926..2964)
                     /label=1P230_D
     primer_bind     complement(2926..2964)
                     /label=1P230_C
     primer_bind     complement(2926..2964)
                     /label=1P230_B
     primer_bind     complement(2926..2964)
                     /label=1P230_A
     primer_bind     complement(2926..2964)
                     /label=1_delP230
     primer_bind     complement(2930..2964)
                     /label=1_insSYXP
     primer_bind     complement(2930..2964)
                     
     primer_bind     complement(2930..2964)
                     /label=1LinkerG_ins1
     primer_bind     complement(2938..2973)
                     /label=1K227_D
     primer_bind     complement(2938..2973)
                     /label=1K227_C
     primer_bind     complement(2938..2973)
                     /label=1K227_B
     primer_bind     complement(2938..2973)
                     /label=1K227_A
     primer_bind     complement(2938..2970)
                     /label=1LinkerS-ins1
     primer_bind     2947..2968
                     /label=2_A231x
     primer_bind     2950..2972
                     /label=2_delP230
     primer_bind     2950..2971
                     /label=2_insSYXP
     primer_bind     2953..2970
                     /label=2LinkerG_ins
     primer_bind     2956..2976
                     /label=2LinkerS-ins
     primer_bind     2959..2980
                     /label=2_K227
     primer_bind     complement(3336..3372)
                     /label=1_AceI94D
     primer_bind     complement(3336..3372)
                     /label=1_AceI94C
     primer_bind     complement(3336..3372)
                     /label=1_AceI94B
     primer_bind     complement(3336..3372)
                     /label=1_AceI94A
     primer_bind     complement(3338..3375)
                     /label=1_AceL93D
     primer_bind     complement(3338..3375)
                     /label=1_AceL93C
     primer_bind     complement(3338..3375)
                     /label=1_AceL93B
     primer_bind     complement(3338..3375)
                     /label=1_AceL93A
     primer_bind     complement(3340..3378)
                     /label=1_AceD92D
     primer_bind     complement(3340..3378)
                     /label=1_AceD92C
     primer_bind     complement(3340..3378)
                     /label=1_AceD92B
     primer_bind     complement(3340..3378)
                     /label=1_AceD92A
     primer_bind     complement(3342..3381)
                     /label=1_AceL91D
     primer_bind     complement(3342..3381)
                     /label=1_AceL91C
     primer_bind     complement(3342..3381)
                     /label=1_AceL91B
     primer_bind     complement(3342..3381)
                     /label=1_AceL91A
     primer_bind     complement(3347..3384)
                     /label=1_AceL90D
     primer_bind     complement(3347..3384)
                     /label=1_AceL90C
     primer_bind     complement(3347..3384)
                     /label=1_AceL90B
     primer_bind     complement(3347..3384)
                     /label=1_AceL90A
     primer_bind     complement(3350..3387)
                     /label=1-AceL89D
     primer_bind     complement(3350..3387)
                     /label=1-AceL89C
     primer_bind     complement(3350..3387)
                     /label=1-AceL89B
     primer_bind     complement(3350..3387)
                     /label=1-AceL89A
     primer_bind     3358..3376
                     /label=2_AceI94x
     primer_bind     3361..3378
                     /label=2_AceL93x
     primer_bind     3364..3381
                     /label=2_AceD92x
     primer_bind     3367..3384
                     /label=2_AceL91x
     primer_bind     3370..3387
                     /label=2_AceL90x
     primer_bind     3373..3390
                     /label=2_AceL89x
     primer_bind     3403..3421
                     /label=2Ace-79X
     primer_bind     complement(3625..3657)
                     /label=2DIO-ANv3-Kv2.1
     primer_bind     complement(3625..3642)
                     /label=1AAV-Syn-Ace_BH1
     protein_bind    complement(3648..3681)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    complement(3732..3765)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3783..4371
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4254..4265)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4379..4603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4605..4745
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4820..5275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5557..5661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5662..6522
                     /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"
     rep_origin      6693..7281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gtcgacattg attattgact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cggggagtcg ctgcgacgct gccttcgccc cgtgccccgc tccgccgccg
      841 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      901 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      961 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
     1021 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg
     1081 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg
     1141 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct
     1201 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc
     1261 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc
     1321 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg
     1381 tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc
     1441 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     1501 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga
     1561 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg
     1621 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc
     1681 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     1741 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     1801 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1861 aattcgatga ctccggaata acttcgtata ggatacttta tacgaagtta tgcagaatgg
     1921 tagctggatt gtagctgcta ttagcaatat gaaacctctt aataacttcg tatagcatac
     1981 attatacgaa gttatggcgc gttagaatcg agtagcctcg gggaaatctg ttgcgcaaga
     2041 gatgaagcta tcaatagatg acatactcct catgtcaatc actccttcag tcctggcagg
     2101 gaggggagcc acgggacttg gcatactgga catctcaagc tcttcgggag gcttagactg
     2161 tggagccatt tcctttgtgt tcaggatagg ctgactttgc acctcgttct cgtagcagaa
     2221 cacgttgatg tcgatctggt ccagggggat gtactcgccc tcgctggtga tcctgctctt
     2281 cttgtacagc tcgtccatgc ccatcacatc ggtaaaggcc ttttgccact ccttgaagtt
     2341 gagctcggtc ttggagtgct tgagctccgt cttacggaac acgtacatcg gctggttctt
     2401 cagatagtta gccgccattg gcttggcaaa ggtgtaggtg gtccgcgcag tgctcctgta
     2461 gcgcttgcca tttccagtgg tgtaactcca cttaaaggta ctgatgatgg ttttgtcgtt
     2521 ggggtaagtc ttcttcgacc tgcaccagtc cgcagcggtc agcgagttgg tcatcacagg
     2581 accgtcagca gggaaaccag tccccttcac ctgggcctct cctttgatgt ggcttccctc
     2641 gtaggtgtag cggtagttaa cagtaaggga ggcaccatct tcaaactgca ttgtgcgatg
     2701 gacttggtat ccggagccat ctaccatggc ggcctggaaa ggcgacatcc cgtcagggta
     2761 gggcaggtac tgatggaagc catacccgat atgagggacc agaatccagg gggagaactg
     2821 gaggtcaccc ttggtggact tcaggtttaa ctcctcataa ccatcatttg gattgccggt
     2881 gccctgaccc accatgtcaa agtccacacc gttgatggag ccaaagatgt gtaactcatg
     2941 tgtcgctggg taggacttga agatagtctc atgggcaatg aggcacccca ttccaaacag
     3001 tggcttagcg agcacatcca ggattcccat gagaatggcc tcattgtcga cggacagcac
     3061 gccactacca gacccgaaag cccacacgac aggataaaac acccagagga ttgacaggaa
     3121 gacgagcaga ctgacatagg agctatggta ctcggggctt ttcttcagat cgtctttcca
     3181 agtggcgtta tacatcccgt atgtcaggac tgcctgcatc acacatccag cgataaagta
     3241 cacccatttg aacttgtgtt catcctcgaa ggcgcccaga ataccaaaca cgatcatgaa
     3301 aatgtctgcg ccgatgaccc aagaaatcat cactccgccc atcttggtca tgacgatgag
     3361 atcgagcagg agcagtggtg tggtcagcac ccagctaata tatcttgcgt agaccacctg
     3421 cctttcacca ttcattgtca ggtcctggta gttcacggcc attgcatagt aagcggtagc
     3481 ggcgatagtg cagatggcaa tgttgatata gtatgcgctc ttcagctctg aaggccgaaa
     3541 gtgcacagcc actccaaaag caatagcagc agcagccata atagcaaaga caatccactg
     3601 tgcaatcatg ccggtctcgg tttccacgtc agccatggtg gcctagcata acttcgtata
     3661 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     3721 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatctcc
     3781 ggaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3841 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3901 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3961 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4021 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4081 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4141 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     4201 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4261 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     4321 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggatcct
     4381 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4441 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4501 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4561 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcaggaac ccctagtgat
     4621 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4681 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4741 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4801 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4861 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4921 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4981 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     5041 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5101 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5161 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5221 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5281 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5341 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5401 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5461 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5521 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5581 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5641 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5701 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5761 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5821 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5881 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5941 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6001 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6061 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6121 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6181 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6241 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6301 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6361 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6421 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6481 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6541 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6601 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6661 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6721 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6781 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6841 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6901 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6961 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7021 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7081 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7141 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7201 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7261 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7321 tgctggcctt ttgctcacat gt
//
