LOCUS       pAAV[FLEXon]-EF1        6764 bp    DNA     circular SYN 01-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..130
                     /label=5' ITR
                     /label=nonstandard type: ITR
                     /note="color: #05696d; direction: RIGHT"
                     /note="/application_notes=Allows rescue of virus from 
                     recombinant plasmid and replication of the viral genome; 
                     this ITR is identical to that of the wild-type AAV2 
                     genome."
                     /note="/description=AAV 5' inverted terminal repeat 
                     (functional equivalent of wild-type 5' ITR)"
                     /note="/official_designation=5' ITR"
                     /note="/uuid=6d343a478711450d8e5c28ceb27e5b84"
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        158..1336
                     /label=EF1A
                     /note="color: #f54c6b; direction: RIGHT"
                     /note="/application_notes=Strong promoter."
                     /note="/description=Human eukaryotic translation elongation
                     factor 1 alpha1 promoter"
                     /note="/full_name=Human eukaryotic translation elongation 
                     factor 1 alpha1 promoter"
                     /note="/official_designation=EF1A"
                     /note="/uuid=69c052aac71a4f49bab5b7ae1f46ace8"
     protein_bind    1336..1360
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1361..1394)
                     /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(1427..1460)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1476..1496
                     /codon_start=1
                     /label=kir2.1-C-term-TS-and-ER-export
                     /translation="NLIFIAE"
     CDS             1497..1691
                     /codon_start=1
                     /label=kv2.1-soma-targeting-signal-peptide
                     /translation="EPGGLWEVCGTADEAVNGAHASHVNHSLCPGRQWGHRAGHAAHLQ
                     LLRGLALRGHLLGVQDGLAL"
     CDS             1692..2378
                     /codon_start=1
                     /label=mScarlet3
                     /translation="AAPRASCGVPGHRSLILLHHCVVLIVAGDVQLSVNVEGSRHLDRL
                     LGLVGGLEISQVPASILQSQGHLDVPLQHHILRVQPLCAGLPAHGPLLHHWAIRREVAS
                     SQLHLVDQGAILQAGVLGHTHCAPILKDHHSLPLEALREALLPVVRDVSWVLDEGPAAI
                     HELGAQDVPAEGQGASLGHLQLGCLGALIGSALSFSLNLKLMAIHAALHVHLEPHELLD
                     HSLSAVH"
     CDS             2379..2438
                     /codon_start=1
                     /label=kir2.1-golgi-trafficking-signal
                     /translation="HIDVNLVQGDVLPFRGDPAL"
     CDS             2448..3380
                     /codon_start=1
                     /label=WiChR_L130F_F240A
                     /translation="PRGIGVILLHRHHHCGSISPSSCQGLLVLLINLLSLPLQLIPGIV
                     LNVQSTSQHEHPKDGLAGNVQEGMDDGKGLLRDHSSGVEAQQP*YRKHGPEAKENEHGS
                     LQVLLGTVRHIPDSFSHLIEKEVEEDAEQGAANGKVDVLLRASHNKAGQGNHQQDSCAA
                     CEGQLIGHIHRKTQIHHNEGAGHNVVTVPVSI**ELRGAVSGKVRGLGHIHQSRQKGNG
                     STQGVQLQGPSLGVRVGSKACAPPAVVPEHHP*AQQDHSREAINGGQQGVVHAHLGARG
                     SLGTGGGASIRVGRLVSGGHHSRGCRAGRH"
     misc_feature    complement(3381..3386)
                     /direction=LEFT
                     /label=Kozak
                     /label=nonstandard type: Miscellaneous
                     /note="color: #6d3ed8; direction: LEFT"
                     /note="/application_notes=Facilitates translation 
                     initiation of ATG start codon downstream of the Kozak 
                     sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=ba4f837cfff042e38e7561244b24cfa6"
     protein_bind    3393..3426
                     /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    3459..3492
                     /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(3493..3517)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3523..3770
                     /label=WPRE3
                     /label=nonstandard type: Miscellaneous
                     /note="color: #946a00; direction: RIGHT"
                     /note="/application_notes=Contained only minimal gamma and 
                     alpha subunits of WPRE. Slightly weaker effective compared 
                     to WPRE."
                     /note="/description=Truncated short version of woodchuck 
                     hepatitis virus posttranscriptional regulatory element (248
                     bp)"
                     /note="/official_designation=WPRE3"
                     /note="/uuid=f2d2a70140634b0189319031f36c6213"
     polyA_signal    3801..4008
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4016..4156
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    complement(4016..4145)
                     /direction=LEFT
                     /label=3' ITR
                     /label=nonstandard type: ITR
                     /note="color: #d84e4e; direction: LEFT"
                     /note="/application_notes=Allows rescue of virus from 
                     recombinant plasmid and replication of the viral genome; 
                     this ITR is identical to that of the wild-type AAV2 
                     genome."
                     /note="/description=AAV 3' inverted terminal repeat"
                     /note="/official_designation=3' ITR"
                     /note="/uuid=a5d684b85c014f76a7b6d8e97945f0fb"
     rep_origin      4231..4686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4968..5072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5073..5141,5142..5933)
                     /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      6104..6692
                     /direction=RIGHT
                     /label=pUC ori
                     /note="color: #db6901; direction: RIGHT"
                     /note="/application_notes=Facilitates plasmid replication 
                     in E. coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=b978ad4ff785460db448caf63b728237"
     repeat_region   join(6754..6764,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgggc tccggtgccc gtcagtgggc
      181 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
      241 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      301 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      361 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
      421 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt
      481 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
      541 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
      601 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
      661 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
      721 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
      781 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
      841 ggggtagtct caagctggcc ggcctgctct ggtgcctggt ctcgcgccgc cgtgtatcgc
      901 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
      961 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
     1021 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
     1081 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
     1141 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     1201 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     1261 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     1321 atttcaggtg tcgtgacaag tttgtacaaa aaagcaggct ataacttcgt ataggatact
     1381 ttatacgaag ttatcctcga gtaccggtca cgcgtacgca agcttgataa cttcgtatag
     1441 catacattat acgaagttat cctagggcca cctcaaacct cattttcata gcagaagaac
     1501 ctggtggcct ctgggaagtc tgtggcacag ctgatgaagc tgtcaatgga gctcatgctt
     1561 ctcatgtcaa tcactccctc tgtcctggca ggcagtgggg ccacagggct gggcatgctg
     1621 ctcatctcca gctcctcagg gggcttgctc tgaggggcca tctccttggt gttcaggatg
     1681 ggttggctct ggctgccccc agagcctcct gtggagtgcc tggccacaga tctctcatac
     1741 tgctccacca ctgtgtagtc ctcattgtgg ctggtgatgt ccagctttct gtcaatgttg
     1801 aaggctccag gcatctggac aggcttcttg gccttgtagg tggtcttgaa atcagccagg
     1861 tacctgcctc catccttcag tctcagggcc atcttgatgt cccccttcag caccacatcc
     1921 tcagggtaca gcctctctgt gctggcctcc cagcccatgg tcctcttctg catcactggg
     1981 ccatcaggag ggaagttgcc tcctctcagc ttcaccttgt agatcagggt gccatcctcc
     2041 aggctggtgt cctgggtcac actcactgtg cccccatcct caaagatcat cactctctcc
     2101 cacttgaagc cctcagggaa gctttgcttc cagtagtcag ggatgtcagc tgggtgcttg
     2161 atgaaggccc tgctgccata catgaactgg gggctcagga tgtcccagct gaagggcagg
     2221 gggcctccct tggtcacctt cagcttggct gtctgggtgc cctcataggg tctgccctct
     2281 ccttctccct caatctcaaa ctcatggcca ttcatgctgc cctccatgtg caccttgaac
     2341 ctcatgaact ccttgatcac agcctcagtg ctgtccatca cattgatgtc aatctggtcc
     2401 agggggatgt actccccttc agaggtgatc ctgctcttgg cagcagcccc agaggcatag
     2461 gggtcatcct cctccacagg catcatcact gtggctccat cagccccagc agctgtcagg
     2521 gtcttctggt tctcctcatc aatctcctgt ctctgcctct tcagctcatc ccaggcatag
     2581 tcctcaatgt gcagtctaca tctcagcatg aacaccccaa agatggtctt gcaggtaatg
     2641 tccaggaagg catggatgat ggcaaaggcc tcctcagaga tcactccagt ggtgtggaag
     2701 cccagcagcc ataataccgg aaacatgggc cagaagccaa agaaaatgaa catggctctc
     2761 tccaggtcct tcttggcaca gtcaggcaca tacctgattc tttttctcac ctcattgaaa
     2821 aagaagtaga agaagatgct gaacagggtg cagccaatgg caaagtagat gtacttctca
     2881 gggcttctca caataaagct ggccagggca atcaccagca ggacagctgt gctgcctgtg
     2941 aaggccagct tatagggcac atccaccgta aaacacagat ccatcacaat gaaggggcag
     3001 gtcacaatgt agtcactgta ccagtatcga tttaatgaga actcagaggt gctgtctctg
     3061 gcaaagtcag aggtcttggc cacatacatc aatcccgtca aaaaggcaat ggcagcactc
     3121 agggtgttca gctccagggg ccttctcttg gggtgagggt gggctccaaa gcctgtgccc
     3181 ctccagctgt agtaccagaa caccacccct gagcccaaca ggatcacagc agagaagcca
     3241 tcaatggtgg ccagcagggt gtagttcatg ctcatctggg ggctaggggc agcctgggca
     3301 caggaggtgg tgcctccatt agggttggga gactggtcag tgggggccac cacagtaggg
     3361 ggtgtagggc aggcagacat ggtggctcta gaataacttc gtataaagta tcctatacga
     3421 agttataggt acctgagctc aacatcgata gcactagtat aacttcgtat aatgtatgct
     3481 atacgaagtt atacccagct ttcttgtaca aagtgggaat tcataatcaa cctctggatt
     3541 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3601 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3661 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     3721 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg gaattcctag
     3781 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3841 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3901 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3961 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagg gccgcaggaa
     4021 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4081 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4141 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4201 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4261 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4321 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4381 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4441 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4501 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4561 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4621 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4681 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4741 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4801 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4861 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4921 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5041 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5101 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5161 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5221 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5281 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5341 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5401 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5461 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5521 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5581 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5641 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5701 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5761 ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag
     5821 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5881 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5941 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6001 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6721 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcag
//
