LOCUS       Exported                7363 bp ds-DNA     circular SYN 08-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-DIO-VARNAM2-Kv2.1PR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7363)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 9, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7363
                     /organism="synthetic DNA construct"
                     /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
                     
     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     1987..2023
                     
     primer_bind     1987..2023
                     /label=1DIO-ANv3-Kv2.1
     primer_bind     2002..2024
                     
     primer_bind     2002..2024
                     /label=2Kv2.1px
     CDS             complement(2006..3657)
                     /codon_start=1
                     /label=VARNAM2-Kv2.1PR
                     /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN
                     IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYISWVLTTPLLLLDLIVMTKMGGVMI
                     SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY
                     HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNEAILMGILDVLAKPLFGMGCLIAH
                     ETIFKSILIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMRIKVIEGGPLPFAF
                     DILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGVVTVTQDTSLEDGE
                     LVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDIALKVDGGGHLHCN
                     FVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVAKYSNLGGGMDELY
                     KKSRITSEGEYIPLDQIDINVFCYENEVQSQPILNTKEMAPQSKPPEELEMSSMPSPVA
                     PLPARTEGVIDMRSMSSIDSFISCATDFPEATR"
     primer_bind     complement(2176..2214)
                     
     primer_bind     complement(2176..2214)
                     /label=1Kv2.1px
     primer_bind     2200..2218
                     /label=2TS/ER_inv
     primer_bind     complement(2261..2280)
                     /label=1VActm-inv
     primer_bind     complement(2261..2280)
                     /label=1Ace-mNeon_inv
     primer_bind     2266..2299
                     /label=2Ace-mRub3
     primer_bind     2281..2301
                     /label=2VActm-inv
     primer_bind     2281..2299
                     /label=2mRUby2_SH1
     primer_bind     complement(2481..2499)
                     /label=1mRub3_164
     primer_bind     2485..2521
                     /label=2mRub3_164A
     primer_bind     complement(2531..2553)
                     /label=1mRub3_146
     primer_bind     2539..2575
                     /label=2mRub3_146A
     primer_bind     complement(2935..2991)
                     /label=1AcRub_228/7
     primer_bind     complement(2937..2991)
                     /label=1AcRub_228/6
     primer_bind     complement(2940..2991)
                     /label=1AcRub_d228/5
     primer_bind     complement(2942..2991)
                     /label=1AcRub_d228/4
     primer_bind     complement(2943..2991)
                     /label=1AcRub_d228/3
     primer_bind     complement(2947..2991)
                     /label=1AcRub_d228/2
     primer_bind     complement(2950..2985)
                     /label=1Ace-mRub3
     primer_bind     complement(2951..2991)
                     /label=1AcRub_d228/1
     primer_bind     complement(2953..2991)
                     /label=1AcRub_ins228
     primer_bind     complement(2953..2991)
                     /label=1ARb_LKins2
     primer_bind     2974..2991
                     /label=2Ace-mNeon_inv
     primer_bind     2977..2999
                     /label=2Ac_lnkr
     primer_bind     complement(3008..3045)
                     /label=1Ace_A256M
     primer_bind     complement(3008..3045)
                     /label=1Ace_210A
     primer_bind     3031..3051
                     /label=2Ace_A256
     primer_bind     3031..3049
                     /label=2Ace_210
     primer_bind     complement(3216..3252)
                     /label=1Ace_141A
     primer_bind     3238..3259
                     /label=2Ace_141
     primer_bind     complement(3384..3432)
                     /label=1Ace_DETC
     primer_bind     complement(3384..3415)
                     /label=1Ace_T125C
     primer_bind     complement(3384..3415)
                     /label=1Ace_125A
     primer_bind     complement(3396..3432)
                     /label=1Ace_121A
     primer_bind     3406..3428
                     /label=2Ace_T125
     primer_bind     3418..3441
                     /label=2Ace_121
     primer_bind     complement(3646..3678)
                     
     protein_bind    complement(3669..3702)
                     /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(3753..3786)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3804..4392
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4275..4286)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4400..4624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4626..4766
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4841..5296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5578..5682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5683..6543
                     /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      6714..7302
                     /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 cttatacagc tcgtccatcc cgccgcccag attagagtac ttggccacgg ccacctctct
     2341 ctgcaccaca tatgtctcgt tatcagactc ctcgatgcgc tccagcctgt ggtccacggc
     2401 gtgcactcct ggcatcttga tattgcccac tgtcttcttg gagcggtatg tggtcacaaa
     2461 gttacagtgc aggtgtcctc ctccatccac cttcagggcg atgtcggtat atcccctcag
     2521 gcctccgtct gcagggtaca tcatctctgt attgggctcc cagcccttgg tcttcttctg
     2581 catcactggg ccattgctag ggaagttcac gcccctcacc ttcacgttat acaccagctc
     2641 gccatcctcc agagaggtgt cctgtgtcac ggtcaccacg ccgccatcct cgtatcttgt
     2701 caccctctcc caggtaaagc cctcggggaa gctctgctta aagaaatcgg ggatgtcggc
     2761 tggatacttg atgaaggtcc tgctgccgta catgaagctg gtggccagga tgtcaaaggc
     2821 gaaaggcagt ggtcctccct cgatcacttt gatcctcatt gtctgcacgc cctcgtatgg
     2881 cctgccctct ccctctccgg tgcacttgaa ctggtgccca ttgacagacc cttccatcac
     2941 gaccttcatc cgcatatttt ccttaatcag gatggacttg aagatagtct catgggcaat
     3001 gaggcacccc attccaaaca gtggcttagc gagcacatcc aggattccca tgagaatggc
     3061 ctcattgtcg acggacagca cgccactacc agacccgaaa gcccacacga caggataaaa
     3121 cacccagagg attgacagga agacgagcag actgacatag gagctatggt actcggggct
     3181 tttcttcaga tcgtctttcc aagtggcgtt atacatcccg tatgtcagga ctgcctgcat
     3241 cacacatcca gcgataaagt acacccattt gaacttgtgt tcatcctcga aggcgcccag
     3301 aataccaaac acgatcatga aaatgtctgc gccgatgacc caagaaatca tcactccgcc
     3361 catcttggtc atgacgatga gatcgagcag gagcagtggt gtggtcagca cccagctaat
     3421 atatcttgcg tagaccacct gcctttcacc attcattgtc aggtcctggt agttcacggc
     3481 cattgcatag taagcggtag cggcgatagt gcagatggca atgttgatat agtatgcgct
     3541 cttcagctct gaaggccgaa agtgcacagc cactccaaaa gcaatagcag cagcagccat
     3601 aatagcaaag acaatccact gtgcaatcat gccggtctcg gtttccacgt cagccatggt
     3661 ggcctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat
     3721 tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta tgctatacga
     3781 agttatgaat tcgatatctc cggaatcaac ctctggatta caaaatttgt gaaagattga
     3841 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3901 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3961 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4021 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4081 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4141 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     4201 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4261 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4321 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4381 ccgcctcccc gcctggatcc tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4621 atggcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4681 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4741 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4801 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5041 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5161 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     5221 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5281 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5341 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5401 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5461 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5521 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6541 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6841 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7081 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
