LOCUS       sequence-431923-        7483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NL1 insert in AAV9 capsid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  AUTHORS   Hanlon KS, Cheng M, Ferrer M, Ryu JR, Lee B, De La Cruz D, Patel N, 
            Espinoza P, Santoscoy MC, Gong Y, Ng C, Nguyen DM, Nammour J, Clark 
            SQ, Heine VM, Sun W, Kozarsky K, Maguire CA
  TITLE     In vivo selection in non-human primates identifies AAV capsids for 
            on-target CSF delivery to spinal cord
  JOURNAL   Molecular Therapy. 2024 Jun 5. doi.org/10.1016/j.ymthe.2024.05.040
  PUBMED    37745398
REFERENCE   2  (bases 1 to 7483)
  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: "Molecular 
            Therapy. 2024 Jun 5. doi.org/10.1016/j.ymthe.2024.05.040"
COMMENT     Sequence Label: NL1
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(441..1232,1233..1301))
                     /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     1064..1083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1302..1406)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1474..1492
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1530..1552)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1652..1671
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1738..2193)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1875..1896)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2087..2106
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2319..2341
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2333..2350
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2334..2350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2357..2376
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2357..2375
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6488..6499
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(6724..6742)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6763..6779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6763..6779)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6776..6798)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6787..6803
                     /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(6811..6817,6818..6835,6836..6841))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6856..6877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6994..7011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7165..7483,1..270))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7245..7264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
       61 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      121 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     1441 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1501 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1561 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1621 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1681 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     1741 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1801 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1861 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1921 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1981 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2041 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2101 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2161 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     2221 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2281 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2341 gacggccagt gaattgtaat acgactcact atagggcgaa ttcgagctcg gtacccctag
     2401 aggtcctgta ttagaggtca cgtgagtgtt ttgcgacatt ttgcgacacc atgtggtcac
     2461 gctgggtatt taagcccgag tgagcacgca gggtctccat tttgaagcgg gaggtttgaa
     2521 cgcgcagcca ccacgccggg gttttacgag attgtgatta aggtccccag cgaccttgac
     2581 gagcatctgc ccggcatttc tgacagcttt gtgaactggg tggccgagaa ggaatgggag
     2641 ttgccgccag attctgacat ggatctgaat ctgattgagc aggcacccct gaccgtggcc
     2701 gagaagctgc agcgcgactt tctgacggaa tggcgccgtg tgagtaaggc cccggaggcc
     2761 cttttctttg tgcaatttga gaagggagag agctacttcc acatgcacgt gctcgtggaa
     2821 accaccgggg tgaaatccat ggttttggga cgtttcctga gtcagattcg cgaaaaactg
     2881 attcagagaa tttaccgcgg gatcgagccg actttgccaa actggttcgc ggtcacaaag
     2941 accagaaatg gcgccggagg cgggaacaag gtggtggatg agtgctacat ccccaattac
     3001 ttgctcccca aaacccagcc tgagctccag tgggcgtgga ctaatatgga acagtattta
     3061 agcgcctgtt tgaatctcac ggagcgtaaa cggttggtgg cgcagcatct gacgcacgtg
     3121 tcgcagacgc aggagcagaa caaagagaat cagaatccca attctgatgc gccggtgatc
     3181 agatcaaaaa cttcagccag gtacatggag ctggtcgggt ggctcgtgga caaggggatt
     3241 acctcggaga agcagtggat ccaggaggac caggcctcat acatctcctt caatgcggcc
     3301 tccaactcgc ggtcccaaat caaggctgcc ttggacaatg cgggaaagat tatgagcctg
     3361 actaaaaccg cccccgacta cctggtgggc cagcagcccg tggaggacat ttccagcaat
     3421 cggatttata aaattttgga actaaacggg tacgatcccc aatatgcggc ttccgtcttt
     3481 ctgggatggg ccacgaaaaa gttcggcaag aggaacacca tctggctgtt tgggcctgca
     3541 actaccggga agaccaacat cgcggaggcc atagcccaca ctgtgccctt ctacgggtgc
     3601 gtaaactgga ccaatgagaa ctttcccttc aacgactgtg tcgacaagat ggtgatctgg
     3661 tgggaggagg ggaagatgac cgccaaggtc gtggagtcgg ccaaagccat tctcggagga
     3721 agcaaggtgc gcgtggacca gaaatgcaag tcctcggccc agatagaccc gactcccgtg
     3781 atcgtcacct ccaacaccaa catgtgcgcc gtgattgacg ggaactcaac gaccttcgaa
     3841 caccagcagc cgttgcaaga ccggatgttc aaatttgaac tcacccgccg tctggatcat
     3901 gactttggga aggtcaccaa gcaggaagtc aaagactttt tccggtgggc aaaggatcac
     3961 gtggttgagg tggagcatga attctacgtc aaaaagggtg gagccaagaa aagacccgcc
     4021 cccagtgacg cagatataag tgagcccaaa cgggtgcgcg agtcagttgc gcagccatcg
     4081 acgtcagacg cggaagcttc gatcaactac gcagacaggt accaaaacaa atgttctcgt
     4141 cacgtgggca tgaatctgat gctgtttccc tgcagacaat gcgagagaat gaatcagaat
     4201 tcaaatatct gcttcactca cggacagaaa gactgtttag agtgctttcc cgtgtcagaa
     4261 tctcaacccg tttctgtcgt caaaaaggcg tatcagaaac tgtgctacat tcatcatatc
     4321 atgggaaagg tgccagacgc ttgcactgcc tgcgatctgg tcaatgtgga tttggatgac
     4381 tgcatctttg aacaataaat gatttaaatc aggtatggct gccgatggtt atcttccaga
     4441 ttggctcgag gacaacctta gtgaaggaat tcgcgagtgg tgggctttga aacctggagc
     4501 ccctcaaccc aaggcaaatc aacaacatca agacaacgct cgaggtcttg tgcttccggg
     4561 ttacaaatac cttggacccg gcaacggact cgacaagggg gagccggtca acgcagcaga
     4621 cgcggcggcc ctcgagcacg acaaggccta cgaccagcag ctcaaggccg gagacaaccc
     4681 gtacctcaag tacaaccacg ccgacgccga gttccaggag cggctcaaag aagatacgtc
     4741 ttttgggggc aacctcgggc gagcagtctt ccaggccaaa aagaggcttc ttgaacctct
     4801 tggtctggtt gaggaagcgg ctaagacggc tcctggaaag aagaggcctg tagagcagtc
     4861 tcctcaggaa ccggactcct ccgcgggtat tggcaaatcg ggtgcacagc ccgctaaaaa
     4921 gagactcaat ttcggtcaga ctggcgacac agagtcagtc ccagaccctc aaccaatcgg
     4981 agaacctccc gcagccccct caggtgtggg atctcttaca atggcttcag gtggtggcgc
     5041 accagtggca gacaataacg aaggtgccga tggagtgggt agttcctcgg gaaattggca
     5101 ttgcgattcc caatggctgg gggacagagt catcaccacc agcacccgaa cctgggccct
     5161 gcccacctac aacaatcacc tctacaagca aatctccaac agcacatctg gaggatcttc
     5221 aaatgacaac gcctacttcg gctacagcac cccctggggg tattttgact tcaacagatt
     5281 ccactgccac ttctcaccac gtgactggca gcgactcatc aacaacaact ggggattccg
     5341 gcctaagcga ctcaacttca agctcttcaa cattcaggtc aaagaggtta cggacaacaa
     5401 tggagtcaag accatcgcca ataaccttac cagcacggtc caggtcttca cggactcaga
     5461 ctatcagctc ccgtacgtgc tcgggtcggc tcacgagggc tgcctcccgc cgttcccagc
     5521 ggacgttttc atgattcctc agtacgggta tctgacgctt aatgatggaa gccaggccgt
     5581 gggtcgttcg tccttttact gcctggaata tttcccgtcg caaatgctaa gaacgggtaa
     5641 caacttccag ttcagctacg agtttgagaa cgtacctttc catagcagct acgctcacag
     5701 ccaaagcctg gaccgactaa tgaatccact catcgaccaa tacttgtact atctctcaaa
     5761 gactattaac ggttctggac agaatcaaca aacgctaaaa ttcagtgtgg ccggacccag
     5821 caacatggct gtccagggaa gaaactacat acctggaccc agctaccgac aacaacgtgt
     5881 ctcaaccact gtgactcaaa acaacaacag cgaatttgct tggcctggag cttcttcttg
     5941 ggctctcaat ggacgtaata gcttgatgaa tcctggacct gctatggcca gccacaaaga
     6001 aggagaggac cgtttctttc ctttgtctgg atctttaatt tttggcaaac aaggaactgg
     6061 aagagacaac gtggatgcgg acaaagtcat gataaccaac gaagaagaaa ttaaaactac
     6121 taacccggta gcaacggagt cctatggaca agtggccaca aaccaccaga gtgcccaact
     6181 gacgacggag gggcggcgtg cacaggcgca gaccggctgg gttcaaaacc aaggaatact
     6241 tccgggtatg gtttggcagg acagagatgt gtacctgcaa ggacccattt gggccaaaat
     6301 tcctcacacg gacggcaact ttcacccttc tccgctgatg ggagggtttg gaatgaagca
     6361 cccgcctcct cagatcctca tcaaaaacac acctgtacct gcggatcctc caacggcctt
     6421 caacaaggac aagctgaact ctttcatcac ccagtattct actggccaag tcagcgtgga
     6481 gatcgagtgg gagctgcaga aggaaaacag caagcgctgg aacccggaga tccagtacac
     6541 ttccaactat tacaagtcta ataatgttga atttgctgtt aatactgaag gtgtatatag
     6601 tgaaccccgc cccattggca ccagatacct gactcgtaat ctgtaattac gtgttaatca
     6661 ataaaccggt tgattcgttt cagttgaact ttggtgtcgc ggccgctcga taagcttttg
     6721 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     6781 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6841 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6901 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6961 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7021 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7081 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aac
//
