LOCUS       sequence-410358-        7526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  production of AAV for cre-dependent expression of Unc5C, tagged with
            3xHA at C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7526)
  AUTHORS   Prigge CL, Dembla M, Sharma A, El-Quessny M, Kozlowski C, Paisley 
            CE, Miltner AM, Johnson TM, Della Santina L, Feller MB, Kay JN
  TITLE     Rejection of inappropriate synaptic partners in mouse retina 
            mediated by transcellular FLRT2-UNC5 signaling.
  JOURNAL   Dev Cell. 2023 Oct 23;58(20):2080-2096.e7. doi: 
            10.1016/j.devcel.2023.07.011. Epub 2023 Aug 8.
  PUBMED    37557174
REFERENCE   2  (bases 1 to 7526)
  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: 
            "10.1016/j.devcel.2023.07.011"; journalName: "Dev Cell"; date: 
            "2023-10-23- 23"; volume: "58"; issue: "20"; pages: "2080-2096.e7"
COMMENT     Sequence Label: pAAV-CMV-flex-Unc5cHA-WPRE-SV40pA
FEATURES             Location/Qualifiers
     source          1..7526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(103..136)
                     /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(187..220)
                     /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)."
     CDS             complement(230..310)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     protein_bind    3121..3154
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3205..3238
                     /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)."
     misc_feature    3245..3833
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3298..3318)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3716..3727)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3876..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3887..3906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3941..3960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4069..4209
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4069..4198
                     /label=AAV2 ITR
     rep_origin      4284..4739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4371..4390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4581..4602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4756..4775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4875..4897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4935..4953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5021..5125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5126..5194,5195..5986)
                     /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(5344..5363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6157..6745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6646..6665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6807..6936
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        6943..7246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aattaagagg taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct
       61 ggagcacctg cctgaaatca ctttttttca ggttgggaat tcataacttc gtatagcata
      121 cattatacga agttattctt tgcaccattc taaagaataa cagtgataat ttctgggtta
      181 aggcaaataa cttcgtatag gatactttat acgaagttat gctagcttag gcatagtcgg
      241 ggacatcata tgggtatgca taatcaggca cgtcgtaggg ataagcgtaa tctggaacat
      301 cgtatgggta accggtatac tgtccttctg ctgccaagga caccactgtc tcatgtcttc
      361 ccatttcttc caggacggct gccagcatgc tcaggtttcc atctgggaag ttctgtgctt
      421 cccaaagatc caggattacg ccagttgggc tcgatttggt ggcaaagtaa ttcaagtacc
      481 tgtccaggtt gagtttatgg gccagcatcc tccagtcatg gcctcttgtt tgaggggcat
      541 ccaggctgct gcatagcttc tgccggatag ggagaggaat gctgaaagca cttggtccgg
      601 tgacagtggt gatggtacta gcagggtcca ggagaggtaa gtcgatgcca gtaggttcct
      661 ctgacacagt acagttgagc tggaagatct gcccttctcc ttcaacctgc cgcacacaga
      721 gtttgcaaac cagttccact gtgtttaggc tgagtctttc cagagtgaag gtgcagtgga
      781 ggtttctttg agagccactc cagatgtggt aaaatggaat ttcctgatac ttagccagca
      841 atttgctctt ccagagggaa tgggcgatgt catgaataga caggcgcagg ttgtggatgc
      901 tgcctttaaa atgaagagcc ttgggttctt ctaggagctg tcctcccatt tgcctctcca
      961 gttgtagaac ttccttcagg gcatcctgtg tgtcatccag gcagtagact ctaatgctgt
     1021 actccaggga agagcagcag aggggcccaa agatggccag tttaagacgc ttggcagctg
     1081 ctttggtggt ggactgccca accagggcat aggtactgag gttctctgtg aggatatggc
     1141 aagcctctgc atccagctga atgtaacagg gggttgtgaa gttctcctcc ccaaccacca
     1201 caacatcctc ccattgtccc tgcactgcct ggtttttgag ctggatcttc cagtcctcgg
     1261 tgctggggtc tgcacagtga tgcagagtga ggatgacagg gcgggtcagc agagctccag
     1321 gaggcccaca gctcaccaca ggggtaagta gggtctgaga gtcttccatg gggggcctca
     1381 tattttcttt cctgtgtaca gtcacataca tttcatagac tctcccctga ggaatggccc
     1441 cagcgggaat cagcaagctt actcctgaat taggaatgat gaggtgaccc ccaagagagt
     1501 tgaaggtacc aaatgctgtg caggatgggt cagtctgtct tgcgaggctc tggttcttca
     1561 ggttaagggc ctcattctct agcaaggact gggtcatctg gggtgacagt ttggatgaga
     1621 actcggcaag gtcatcctga ggagtgacag cacctgagct gttgtacact ttgattttca
     1681 agttgggtag tgggtccaga attggagagt tggtcattgg gattttgtct gagacatcat
     1741 gcagagcata gacaggtccc ctgtacatgg ctgcagctga ggtgaggtca ggggggacag
     1801 ccaggagatc ttgtctggca gccttgatgt tcacaggctg aaagccgcca ttgagtgctg
     1861 aggagtcaat gatgtcagac tcaaagtcac ggtggttctt ccgatacaca aacagggcca
     1921 ccacaacagt gatcgccaga cagactgtta cagcgatcac aatccccacg tagagagcca
     1981 catcatctga gtcaggagca gcctgcatgc acagcccatc agtgcagttc ttggattgga
     2041 ggaccaggcc atcacagtcc ttacccccgt tcttgggggc tggtgctgta cactccctcc
     2101 tgcgccagtg ggtgcattca gtcccacagg ttgaccattt gctccatgaa gtccacctac
     2161 catccactgg acataacgta gtgcatgcta ttttctgcac actctgcccc tcacagaagg
     2221 ccccaccatt gagtggggct gggttggtgc aggttcttgt gcgtttctga tatcctcgcc
     2281 cacagcggct gttacacaca gaccactctg tccaggtgga ccagccacca ttaacataca
     2341 cgatgacagt ggctgtggtg ctttttctct tggcaacaat atttttggca acacaggtat
     2401 aatttgctgt atctgagagt cgggcttgct tgatgatcag gttgtgatcg atagtaatat
     2461 aaaagttccg atcttcagca ggatcaatta tgtcttcatt ctttagccat tctacctcag
     2521 ccactgggat cccttcaggt ggccgacact ggagtaagac ttcctgctcc aaggacactt
     2581 cctttcccaa gggttcctgc tcgaatgtct tccgcagata cgcaatgcgc acgtatgcct
     2641 tccgactctt cgtagtgcct gctgagctcc aggccacaca ctggcaccag taatcttcag
     2701 gcccaaacag ttcctccacc tgctggcgtg aaatctcaat gctcacttct ctcacaatta
     2761 gaccagaggt ttcatctact ctctcgtcta ctacgtggtc cttctgatga acccactcgc
     2821 tgttgcactt gaagtagatc tgggtggcag ggctggcttt acaatacagg ttcacaggct
     2881 tgttcttcac aatgtaagct tcctcgggct caatgaggaa gtgtggcaat ggctcaggtg
     2941 ggtcagatgg aaaggtttct gggagttcgt gaaaaaattc atcatcttga gcggcggagc
     3001 cggtgccact ggcgcttagc agggccaggg caggtaacac aagcatctgc agcaagtatc
     3061 ctagtcccag tccgcagcgg gccgctgtcg ccctcagacc tttcctcatg gtggcgcgcc
     3121 ataacttcgt ataatgtatg ctatacgaag ttattaagag gtttcatatt gctaatagca
     3181 gctacaatcc agctaccatt ctgcataact tcgtataaag tatcctatac gaagttattc
     3241 cggaaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3301 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3361 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3421 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3481 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3541 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3601 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3661 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3721 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3781 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3841 ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4021 tatgtttcag gttcaggggg agatgtggga ggttttttaa agggccgcag gaacccctag
     4081 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4141 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4201 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4261 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4621 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4681 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4741 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4801 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4861 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4921 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4981 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5761 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5821 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5881 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5941 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6781 cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga
     6841 ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     6901 gagagggagt ggccaactcc atcactaggg gttcctgtcg accgttacat aacttacggt
     6961 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     7021 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7081 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     7141 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     7201 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
     7261 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
     7321 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggs gsgsgcsggg
     7381 gggggggggg gggggggggg gggggggsgg ggcgaggcgg agaggtgcgg cggcagccaa
     7441 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
     7501 taaaaagcga agcgcgcggc gggcgg
//
