LOCUS       sequence-465217-        8543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-standard AAV2 rep-AAV-PHP.eB cap plasmid with AAV cap expression
            controlled by a tTA-TRE amplification system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8543)
  AUTHORS   Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, 
            Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V
  TITLE     Engineered AAVs for efficient noninvasive gene delivery to the 
            central and peripheral nervous systems.
  JOURNAL   Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 
            2017 Jun 26.
  PUBMED    28671695
REFERENCE   2  (bases 1 to 8543)
  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/nn.4593"; 
            journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue: 
            "8"; pages: "1172-1179"
COMMENT     Sequence Label: pUCmini-iCAP-PHP.eB
FEATURES             Location/Qualifiers
     source          1..8543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1145..1156)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    3873..3891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3910..3928
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3947..3965
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3984..4002
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4021..4039
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4058..4076
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4095..4113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4132..4150
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    4212..4436
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4425..4442
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        4462..4480
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4462..4479
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4519..4629)
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     primer_bind     complement(7218..7237)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7219..7237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7247..7264)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7247..7263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7256..7278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        7408..7512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7513..7581,7582..8373)
                     /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     complement(7731..7750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttccgat
      661 tcggtccgca aaagctggag ctccaccgcg gtggcggccg ctctagtcga gcggccgcca
      721 gtgtgatgga tcgatagttc ctaggacatg tgatatctgc agaattcggc ttggcggctg
      781 cgcgttcaaa cctcccgctt caaaatggag accctgcgtg ctcactcggg cttaaatacc
      841 cagcgtgacc acatggtgtc gcaaaatgtc gcaaaacact cacgtgacct ctaatacagg
      901 acctctagtc ctgcaggttt aaacgaattg cccttcgcag agaccaaagt tcaactgaaa
      961 cgaattaaac ggtttattga ttaacaagca agtcgactta cagattacga gtcaggtatc
     1021 tggtgccaat ggggcggggt tcactatata caccttcagt attaacagca aattcaacat
     1081 tattagactt gtaatagttg gaagtgtact ggatctccgg gttccagcgc ttgctgtttt
     1141 ccttctgcag ctcccactcg atctccacgc tgacttgacc agtagaatac tgggtgatga
     1201 aagagttcag cttgtccttg ttgaaggccg ttggaggatc cgcaggtaca ggtgtgtttt
     1261 tgatgaggat ctgaggaggc gggtgcttca ttccaaaccc tcccatcagc ggagaagggt
     1321 gaaagttgcc gtccgtgtga ggaattttgg cccaaatggg tccttgcagg tacacatctc
     1381 tgtcctgcca aaccataccc ggaagtattc cttggttttg aacccaaccg gtctgcgcct
     1441 gtgccttaaa aggcaccgcc aaagtcccat cactctggtg gtttgtggcc acttgtccat
     1501 aggactccgt tgctaccggg ttagtagttt taatttcttc ttcgttggtt atcatgactt
     1561 tgtccgcatc cacgttgtct ctgccagtac cttgtttgcc aaaaattaaa gatccagaca
     1621 aaggaaagaa acggtcctct ccttctttgt gagaggccat agcaggtcca ggattcatca
     1681 agctattacg tccattgaga gcccaagaag aagctccagg ccaagcaaat tcgctgttgt
     1741 tgttttgagt cacagtggtt gagacacgtt gttgtcggta gctgggtcca ggtatgtagt
     1801 ttcttccctg gacagccatg ttgctgggtc cggccacact gaattttagc gtttgttgat
     1861 tctgtccaga accgttaata gttctagaga gatagtacaa gtattggtcg atgagtggat
     1921 tcattagtcg gtccaggctt tggctgtgag cgtagctgct atggaaaggt acgttctcaa
     1981 actcgtagct gaactggaag ttgttacccg ttcttagcat ttgcgacggg aaatattcca
     2041 ggcagtaaaa ggacgaacga cccacggcct ggcttccatc attaagcgtc agatacccgt
     2101 actgaggaat catgaaaacg tccgctggga acggcgggag gcagccctcg tgagccgacc
     2161 cgagcacgta cgggagctga tagtctgagt ccgtgaagac ctggaccgtg ctggtaaggt
     2221 tattggcgat ggtcttgact ccattgttgt ccgtaacctc tttgacctga atgttaaaga
     2281 gcttgaagtt gagtcgctta ggccggaatc cccagttgtt gttgatgagt cgctgccagt
     2341 cacgtggtga gaagtggcag tggaatctgt tgaagtcaaa atacccccag ggggtgctgt
     2401 agccgaagta ggcgttgtca tttgaagatc ctccagatgt gctgttggag atttgcttgt
     2461 agaggtgatt gttgtaggtg ggcagggccc aggttcgggt gctggtggtg atgactctgt
     2521 cccccagcca ttgggaatcg caatgccaat ttcccgagga actacccact ccatcggcac
     2581 cttcgttatt gtctgccact ggtgcgccac cacctgaagc cattgtaaga gatcccacac
     2641 ctgagggggc tgcgggaggt tctccgattg gttgagggtc tgggactgac tctgtgtcgc
     2701 cagtctgacc gaaattgagt ctctttttag cgggctgtgc acccgatttg ccaatacccg
     2761 cggaggagtc cggttcctga ggagactgct ctacaggcct cttctttcca ggagccgtct
     2821 tagccgcttc ctcaaccaga ccaagaggtt caagaagcct ctttttggcc tggaagactg
     2881 ctcgcccgag gttgccccca aaagacgtat cttctttgag ccgctcctgg aactcggcgt
     2941 cggcgtggtt gtacttgagg tacgggttgt ctccggcctt gagctgctgg tcgtaggctt
     3001 tgtcgtgctc gagggccgcc gcgtctgctg cgttgaccgg ctcccccttg tcgagtccgt
     3061 tgccgggtcc aaggtatttg taacccggaa gcacaagacc tctagcgttg tcttgatgtt
     3121 gttgatttgc cttgggttga ggggctccag gtttcaaagc ccaccactcg cgaattcctt
     3181 cactaaggtt gtcctcgagc caatctggaa gataaccatc ggcagccata cctggcttaa
     3241 gtcatttatt gttcaaagat gcagtcatcc aaatccacat tgaccagatc gcaggcagtg
     3301 caagcgtctg gcacctttcc catgatatga tgaatgtagc acagtttctg atacgccttt
     3361 ttgacgacag aaacgggttg agattctgac acgggaaagc actctaaaca gtctttctgt
     3421 ccgtgagtga agcagatatt tgagttctga ttcattctct cgcattgtct gcagggaaac
     3481 agaatcagat taatgcccac gtgacgagaa cacttgtttt ggtacctgtc tgcgtagttg
     3541 atcgaagcta gcggtcgcag aggagcggga tcaacagtca cgtctgaact gcgaggcgtc
     3601 tcgggaacaa atgagagcct ggcccgcttc tccagacttt tatagctagt attggtgacg
     3661 tcacccagtg ggcgttttag agatttctcc gcccctttag ttcccgccaa ttccctggga
     3721 actttaaact cgtgagtcac cggcacctga ttgacctttg cccaagcaaa aaagtccttg
     3781 acttcctgct tagtaatctt gccaaaatct ggcgggagcc gcttagtcag ttcaaatttg
     3841 aacagatctc tctgcttata tagggggtac cttctctatc actgataggg agctcgagac
     3901 gtgtcgactt ctctatcact gatagggaga cgttctagac gtaccttctc tatcactgat
     3961 agggagctcg agacgtgtcg acttctctat cactgatagg gagacgttct agacgtacct
     4021 tctctatcac tgatagggag ctcgagacgt gtcgacttct ctatcactga tagggagacg
     4081 ttctagacgt accttctcta tcactgatag ggagctcgag acgtgtcgac ttctctatca
     4141 ctgataggga gacgtcaggt ggcacttgat agcttgattc gagccccagc tggttctttc
     4201 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
     4261 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     4321 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     4381 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     4441 ggctgatcag cgagctctag catttaggtg acactataga atagggccct ctagatcgaa
     4501 ttcatgcatt tacccgggga gcatgtcaag gtcaaaatcg tcaagagcgt cagcaggcag
     4561 catatcaagg tcaaagtcgt caagggcatc ggctgggagc atgtctaagt caaaatcgtc
     4621 aagggcgtcg gccggcccgc cgctttcgca ctttagctgt ttctccaggc cacatatgat
     4681 tagttccagg ccgaaaagga aggcaggttc ggctccctgc cggtcgaaca gctcaattgc
     4741 ttgtctcaga agtgggggca tagaatcggt ggtaggtgtc tctctttcct cttttgctac
     4801 ttgatgctcc tgttcctcca atacgcagcc cagtgtaaag tggcccacgg cggacagagc
     4861 gtacagtgcg ttctccaggg agaagccttg ctgacacagg aacgcgagct gattttccag
     4921 ggtttcgtac tgtttctctg ttgggcgggt gccgagatgc actttagccc cgtcgcgatg
     4981 tgagaggaga gcacagcgga atgacttggc gttgttccgc agaaagtctt gccatgactc
     5041 gccttccagg gggcagaagt gggtatgatg cctgtccagc atctcgatgg ccagggcatc
     5101 gagcagggcc cgcttgttct tcacgtgcca gtacagggta ggctgctcaa ctcccagctt
     5161 ttgagcgagt ttccttgtcg tcaggccttc gataccgact tcattgagta attccagagc
     5221 gccgtttatg actttagatt tatccagccg actcatacct ggtttaagtc atttattgtt
     5281 cagaaacaca gtcatccaag tccacattga ccaggtcgca agcagtgcaa gcgtctggca
     5341 cctttcccat gatgtgatga atgtagcaca gtttctgata cgcctttttg acgacagaaa
     5401 cgggttgaga ttctgacacg ggaaagcact ctaaacagtc tttgacaccg tgagtgaagc
     5461 agatatttga attctgattc agtctctcgc attgtctgca gggaaacagc atcagattca
     5521 tgcccacgtg acgagaacat ttgttttggt acctgtccgc gtagttgatc gaagcttccg
     5581 cgtctgacgt cgatggctgc gcaactgact cgcgcacccg tttgggctca cttatatctg
     5641 cgtcactggg ggcgggtctt ttcttggctc cacccttttt gacgtagaat tcatgctcca
     5701 cctcaaccac gtgatccttt gcccaccgga aaaagtcttt gacttcctgc ttggtgacct
     5761 tcccaaagtc atgatccaga cggcgggtga gttcaaattt gaacatccgg tcttgcaacg
     5821 gctgctggtg ttcgaaggtc gttgagttcc cgtcaatcac ggcgcacata ttggtgttgg
     5881 aggtgacgat cacgggagtc gggtctatct gggccgagga cttgcatttc tggtccacgc
     5941 gcaccttgct tcctccgaga atggctttgg ccgactccac gaccttggcg gtcatcttcc
     6001 cctcctccca ccagatcacc atcttgtcca cacagtcgtt gaagggaaag ttctcattgg
     6061 tccagtttac gcacccgtag aagggcacag tgtgggctat ggcctccgcg atgttggtct
     6121 tcccggtagt tgcaggccca aacagccaga tggtgttcct cttgccgaac tttttcgtgg
     6181 cccatcccag aaagacggaa gccgcatatt ggggatcgta cccgtttagt tccaaaattt
     6241 tataaatccg attgctggaa atgtcctcca cgggctgctg gcccaccagg tagtcggggg
     6301 cggttttagt caggctcata atctttcccg cattgtccaa ggcagccttg atttgggacc
     6361 gcgagttgga ggccgcattg aaggagatgt atgaggcctg gtcctcctgg atccactgct
     6421 tctccgaggt aatccccttg tccacgagcc acccgaccag ctccatgtac ctggctgaag
     6481 tttttgatct gatcaccggc gcatcagaat tgggattctg attctctttg ttctgctcct
     6541 gcgtctgcga cacgtgcgtc agatgctgcg ccaccaaccg tttacgctcc gtgagattca
     6601 aacaggcgct taaatactgt tccatattag tccacgccca ctggagctca ggctgggttt
     6661 tggggagcaa gtaattgggg atgtagcact catccaccac cttgttcccg cctccggcgc
     6721 catttctggt ctttgtgacc gcgaaccagt ttggcaaagt cggctcgatc ccgcggtaaa
     6781 ttctctgaat cagtttttcg cgaatctgac tcaggaaacg tcccaaaacc atggatttca
     6841 ccccggtggt ttccacgagc acgtgcatgt ggaagtagct ctctcccttc tcaaattgca
     6901 caaagaaaag agcctccggg gccttactca cacggcgcca ttccgtcaga aagtcgcgct
     6961 gcagcttctc ggccacggtc aggggtgcct gctcaatcag attcagatcc atgtcagaat
     7021 ctggcggcaa ctcccattcc ttctcggcca cccagttcac aaagctgtca gaaatgccgg
     7081 gcagatgctc gtcaaggtcg ctggggacct taatcacaat ctcgtaaaac cccggcatgg
     7141 cggctgcgcg ttcaaacctc ccgcttcaaa atggagaccc tgcgagctcc cccgataccg
     7201 tcgatgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     7261 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     7321 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgacgcgtat cattgggacg
     7381 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     7441 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7501 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7561 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7621 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7681 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7741 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7801 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7861 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7921 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7981 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     8041 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     8101 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     8161 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     8221 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     8281 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     8341 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     8401 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     8461 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8521 gtagaaaaga tcaaaggatc ttc
//
