LOCUS       sequence-135850-        8825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3A protein (truncated at cleavage site P3; ie, 
            long and missing exon 3), C-terminally fused to alkaline phosphatase
            + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8825)
  AUTHORS   Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray 
            TA, Kay JN, Wojtowicz WM
  TITLE     An extracellular biochemical screen reveals that FLRTs and Unc5s 
            mediate neuronal subtype recognition in the retina.
  JOURNAL   Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
  PUBMED    26633812
REFERENCE   2  (bases 1 to 8825)
  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: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Sema3a(L, del3)-AP-His
FEATURES             Location/Qualifiers
     source          1..8825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1573
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     regulatory      1604..1613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5405..5422
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5448..5569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5485..5504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5539..5558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5607..5624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5778..6366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5858..5877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6537..7328,7329..7397))
                     /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     7160..7179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7398..7502)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7570..7588
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7626..7648)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7650..7670)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7655..7985
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7836..7971
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7898..7917
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8122..8141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8208..8663)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8345..8366)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8557..8576
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8789..8811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8803..8820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8804..8820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
      781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
      841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
      901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
      961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
     1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
     1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
     1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
     1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
     1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
     1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
     1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
     1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
     1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
     1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tgggctggtt
     1621 cactgggatt gcctgtcttt tctggggtgt attacttaca gccagagcaa actatgcaaa
     1681 cggaaagaac aatgtgccaa gactgaaatt atcgtacaaa gaaatgttgg aatccaacaa
     1741 tgtgatcact tttaatggct tggccaacag ctccagttac cacaccttcc ttctggatga
     1801 agaacggagt agactatatg ttggagcaaa agatcatata ttttcattca acttggtgaa
     1861 cattaaagat tttcaaaaga ttgtgtggcc agtatcttac acaaggagag atgaatgcaa
     1921 atgggctgga aaagatatcc tgaaagaatg tgccaatttc atcaaggtcc tggaggctta
     1981 taatcagact cacttgtatg cctgtggaac tggggctttc catccaatct gcacctatat
     2041 tgaagttgga catcatcctg aggacaacat ttttaagctg caggactcac attttgaaaa
     2101 cggtcgtggg aagagccctt atgatcccaa actactgact gcctctcttc taatagacgg
     2161 tgagttgtac tctggaactg ctgcggactt catgggacgg gacttcgcta tcttcagaac
     2221 actggggcac catcacccca tcaggacgga gcagcatgac tcccggtggc tcaatgatcc
     2281 tagattcatc agtgcccatc tcatcccaga gagtgacaac cctgaagatg acaaagtata
     2341 ttttttcttc cgagaaaatg caatagacgg agaacactct ggaaaagcca ctcatgctag
     2401 aataggtcag atatgcaaga atgactttgg tggacacaga agtcttgtga ataaatggac
     2461 aacattccta aaagcacgcc tgatttgctc tgtgcccggt cccaatggca ttgacaccca
     2521 ttttgatgaa ttgcaggatg tattcctaat gaactctaaa gatcctaaaa atccgatcgt
     2581 ctatggagtg ttcacaacat caagcaacat ctttaaggga tctgctgtgt gcatgtacag
     2641 catgagtgat gtaagaaggg tgttccttgg tccatatgct cacagagatg gtcccaacta
     2701 tcagtgggtg ccttaccaag gaagagtccc ttatccacgg ccaggaactt gtcccagtaa
     2761 aacatttggc ggatttgact ccacaaagga ccttcctgat gatgtcataa cttttgcaag
     2821 aagtcatcca gccatgtaca acccagtgtt tcctataaat aatcgcccga tcatgatcaa
     2881 aacagatgta aattatcagt tcacacaaat tgttgtagac cgagtggatg cagaagatgg
     2941 ccagtatgat gttatgttca tcggaacaga tgttggaacc gttcttaaag tggtttcagt
     3001 ccccaaggag acttggcatg acctagaaga agttcttctg gaagaaatga ccgtcttccg
     3061 ggaaccaaca actatttcgg caatggagct ttctactaaa cagcaacagc tgtacattgg
     3121 ctcaactgcg ggagtggcac agcttcctct acaccgctgt gacatctatg gcaaagcctg
     3181 tgcagaatgc tgcctcgctc gggaccctta ctgtgcctgg gatgggtcct catgctcacg
     3241 ctattttcct actgcaaagg cccgcacaag agcacaagat ataaggaatg gagacccact
     3301 gactcactgc tctgacttgc agcaccatga taatcatcat gggcccagcc ttgaagagag
     3361 aatcatctat ggagtggaaa acagtagtac attcttggaa tgcagtccga agtcacagag
     3421 agccttggta tattggcaat ttcagaggag aaatgaagat cgaaaagagg agatcagaat
     3481 gggtgatcat atcatcagga cagaacaagg gctcctgctc cgtagcctgc agaagaagga
     3541 ttcaggcaat tacctgtgtc acgctgtgga acacggattc atgcaaactc ttcttaaggt
     3601 aaccctggaa gtcattgaca cagaacattt ggaagaactt cttcataaag atgacgatgg
     3661 agatggctct aagataaaag aaatgtcgag cagcatgacg cccagccaga aagtctggta
     3721 ccgagacttc atgcagctca ttaaccaccc caacctgaac acgatggatg agttctgtga
     3781 acaagtgtgg aaaagggacc gaaagcaacg ccgacaaagg ccggggcact ctcaagggag
     3841 cagcaacaag tggaagcaca tgcaagagag caagaaaggt agaaacaggc gtacgcacga
     3901 gtttgagcgg gcacccggcg cgcctggatc ttccggaatc atcccagttg aggaggagaa
     3961 cccggacttc tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc
     4021 tgcacagaca gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac
     4081 ggtgacagct gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc
     4141 cctggccatg gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca
     4201 tgtgccagac agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca
     4261 gaccattggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga
     4321 ggtcatctcc gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac
     4381 cacacgagtg cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg
     4441 gtactcggac gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac
     4501 gcagctcatc tccaacatgg acattgacgt gatcctaggt gggggccgaa agtacatgtt
     4561 tcgcatggga accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct
     4621 ggacgggaag aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg
     4681 gaaccgcact gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct
     4741 ctttgagcct ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct
     4801 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct
     4861 cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact
     4921 gactgagacg atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga
     4981 ggacacgctg agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc
     5041 cctgcgaggg agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta
     5101 cacggtcctc ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga
     5161 tgttaccgag agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga
     5221 cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct
     5281 ggttcacggc gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct
     5341 ggagccctac accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc
     5401 gggtcatcat caccatcacc attaatctag agatcttatt aaagcagaac ttgtttattg
     5461 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5521 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggt
     5581 cgactctagc tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5641 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5701 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6781 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6841 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6901 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6961 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7021 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7081 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7141 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7201 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7261 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7321 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7381 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7441 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7501 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7561 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     7621 gaaaacctct gacacatgca gctcccggag gaatgtgtgt cagttagggt gtggaaagtc
     7681 cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     7741 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     7801 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     7861 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     7921 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     7981 gcaaaaagct cccgggagct tgtatatcca ttttcgtccc ggagacggtc acagcttgtc
     8041 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     8101 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     8161 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     8221 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     8281 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     8341 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     8401 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     8461 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     8521 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     8581 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     8641 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
     8701 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     8761 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     8821 gaatt
//
