LOCUS       sequence-443194-        7360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging plasmid expressing Rep/Cap genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7360)
  AUTHORS   Zhang Z, Su J, Tang J, Chung L, Page JC, Winter CC, Liu Y, Kegeles 
            E, Conti S, Zhang Y, Biundo J, Chalif JI, Hua CY, Yang Z, Yao X, 
            Yang Y, Chen S, Schwab JM, Wang KH, Chen C, Prerau MJ, He Z
  TITLE     Spinal projecting neurons in rostral ventromedial medulla 
            co-regulate motor and sympathetic tone.
  JOURNAL   Cell. 2024 May 3:S0092-8674(24)00447-1. doi: 
            10.1016/j.cell.2024.04.022.
  PUBMED    38733990
REFERENCE   2  (bases 1 to 7360)
  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"; journalName: "Cell. 2024 
            May 3:S0092-8674(24)00447-1. doi: 10.1016/j.cell.2024.04.022."
COMMENT     Sequence Label: AAV9retro
FEATURES             Location/Qualifiers
     source          1..7360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3845..3856
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4308..4328)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4308..4326)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4360..4382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4371..4387
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4395..4401,4402..4419,4420..4425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4440..4461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4578..4595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4749..5337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4829..4848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5508..6299,6300..6368))
                     /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     6131..6150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6369..6473)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6499..6954)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6636..6657)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6848..6867
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7081..7103
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7095..7112
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7096..7112
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7122..7141
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7122..7140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7169..7185
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7170..7186
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag
       61 cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggctct tttctttgtg
      121 caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac caccggggtg
      181 aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat tcagagaatt
      241 taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac cagaaatggc
      301 gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt gctccccaaa
      361 acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag cgcctgtttg
      421 aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc gcagacgcag
      481 gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag atcaaaaact
      541 tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac ctcggagaag
      601 cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc caactcgcgg
      661 tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac taaaaccgcc
      721 cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg gatttataaa
      781 attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct gggatgggcc
      841 acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac taccgggaag
      901 accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt aaactggacc
      961 aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg ggaggagggg
     1021 aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag caaggtgcgc
     1081 gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat cgtcacctcc
     1141 aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca ccagcagccg
     1201 ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga ctttgggaag
     1261 gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt ggttgaggtg
     1321 gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc cagtgacgca
     1381 gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac gtcagacgcg
     1441 gaagcttcga tcaactacgc ggacaggtac caaaacaaat gttctcgtca cgtgggcatg
     1501 aatctgatgc tgtttccctg cagacaatgc gagagactga atcagaattc aaatatctgc
     1561 ttcactcacg gtgtcaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt
     1621 tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcacatcat gggaaaggtg
     1681 ccagacgctt gcactgcttg cgacctggtc aatgtggact tggatgactg tgtttctgaa
     1741 caataaatga cttaaaccag gtatggctgc cgatggttat cttccagatt ggctcgagga
     1801 caaccttagt gaaggaattc gcgagtggtg ggctttgaaa cctggagccc ctcaacccaa
     1861 ggcaaatcaa caacatcaag acaacgctcg aggtcttgtg cttccgggtt acaaatacct
     1921 tggacccggc aacggactcg acaaggggga gccggtcaac gcagcagacg cggcggccct
     1981 cgagcacgac aaggcctacg accagcagct caaggccgga gacaacccgt acctcaagta
     2041 caaccacgcc gacgccgagt tccaggagcg gctcaaagaa gatacgtctt ttgggggcaa
     2101 cctcgggcga gcagtcttcc aggccaaaaa gaggcttctt gaacctcttg gtctggttga
     2161 ggaagcggct aagacggctc ctggaaagaa gaggcctgta gagcagtctc ctcaggaacc
     2221 ggactcctcc gcgggtattg gcaaatcggg tgcacagccc gctaaaaaga gactcaattt
     2281 cggtcagact ggcgacacag agtcagtccc agaccctcaa ccaatcggag aacctcccgc
     2341 agccccctca ggtgtgggat ctcttacaat ggcttcaggt ggtggcgcac cagtggcaga
     2401 caataacgaa ggtgccgatg gagtgggtag ttcctcggga aattggcatt gcgattccca
     2461 atggctgggg gacagagtca tcaccaccag cacccgaacc tgggccctgc ccacctacaa
     2521 caatcacctc tacaagcaaa tctccaacag cacatctgga ggatcttcaa atgacaacgc
     2581 ctacttcggc tacagcaccc cctgggggta ttttgacttc aacagattcc actgccactt
     2641 ctcaccacgt gactggcagc gactcatcaa caacaactgg ggattccggc ctaagcgact
     2701 caacttcaag ctcttcaaca ttcaggtcaa agaggttacg gacaacaatg gagtcaagac
     2761 catcgccaat aaccttacca gcacggtcca ggtcttcacg gactcagact atcagctccc
     2821 gtacgtgctc gggtcggctc acgagggctg cctcccgccg ttcccagcgg acgttttcat
     2881 gattcctcag tacgggtatc tgacgcttaa tgatggaagc caggccgtgg gtcgttcgtc
     2941 cttttactgc ctggaatatt tcccgtcgca aatgctaaga acgggtaaca acttccagtt
     3001 cagctacgag tttgagaacg tacctttcca tagcagctac gctcacagcc aaagcctgga
     3061 ccgactaatg aatccactca tcgaccaata cttgtactat ctctcaaaga ctattaacgg
     3121 ttctggacag aatcaacaaa cgctaaaatt cagtgtggcc ggacccagca acatggctgt
     3181 ccagggaaga aactacatac ctggacccag ctaccgacaa caacgtgtct caaccactgt
     3241 gactcaaaac aacaacagcg aatttgcttg gcctggagct tcttcttggg ctctcaatgg
     3301 acgtaatagc ttgatgaatc ctggacctgc tatggccagc cacaaagaag gagaggaccg
     3361 tttctttcct ttgtctggat ctttaatttt tggcaaacaa ggaactggaa gagacaacgt
     3421 ggatgcggac aaagtcatga taaccaacga agaagaaatt aaaactacta acccggtagc
     3481 aacggagtcc tatggacaag tggccacaaa ccaccagagt gcccaactag cagaccaaga
     3541 ctacacaaaa actgctgcac aggcgcagac cggctgggtt caaaaccaag gaatacttcc
     3601 gggtatggtt tggcaggaca gagatgtgta cctgcaagga cccatttggg ccaaaattcc
     3661 tcacacggac ggcaactttc acccttctcc gctgatggga gggtttggaa tgaagcaccc
     3721 gcctcctcag atcctcatca aaaacacacc tgtacctgcg gatcctccaa cggccttcaa
     3781 caaggacaag ctgaactctt tcatcaccca gtattctact ggccaagtca gcgtggagat
     3841 cgagtgggag ctgcagaagg aaaacagcaa gcgctggaac ccggagatcc agtacacttc
     3901 caactattac aagtctaata atgttgaatt tgctgttaat actgaaggtg tatatagtga
     3961 accccgcccc attggcacca gatacctgac tcgtaatctg taattgcttg ttaatcaata
     4021 aaccgtttaa ttcgtttcag ttgaactttg gtctctgcga agggcgaatt cgtttaaacc
     4081 tgcaggacta gaggtcctgt attagaggtc acgtgagtgt tttgcgacat tttgcgacac
     4141 catgtggtca cgctgggtat ttaagcccga gtgagcacgc agggtctcca ttttgaagcg
     4201 ggaggtttga acgcgcagcc gccaagccga attctgcaga tatccatcac actggcggcc
     4261 gctcgactag agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
     4321 ttaattgcgc gcttggcgta atcatggtta tagctgtttc ctgtgtgaaa ttgttatccg
     4381 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4441 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4501 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4561 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4621 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4681 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4741 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4801 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4861 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4921 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4981 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5041 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5101 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5161 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5221 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5281 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5341 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5401 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5461 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5521 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5581 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5641 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5701 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5761 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5821 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5881 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5941 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6001 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6061 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6121 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6181 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6241 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6301 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6361 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6421 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6481 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6541 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6601 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6661 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6721 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6781 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6841 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6901 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     6961 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7021 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7081 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     7141 gcgaattggg taccgggccc cccctcgatc gaggtcgacg gtatcggggg agctcgcagg
     7201 gtctccattt tgaagcggga ggtttgaacg cgcagccgcc atgccggggt tttacgagat
     7261 tgtgattaag gtccccagcg accttgacga gcatctgccc ggcatttctg acagctttgt
     7321 gaactgggtg gccgagaagg aatgggagtt gccgccagat
//
