LOCUS       sequence-135627-        8795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3D protein (truncated at cleavage site P3; ie, 
            long), 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 8795)
  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 8795)
  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: Sema3d(L)-AP-His
FEATURES             Location/Qualifiers
     source          1..8795
                     /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)"
     CDS             5375..5392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5418..5539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5455..5474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5509..5528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5577..5594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5748..6336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5828..5847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6507..7298,7299..7367))
                     /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     7130..7149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7368..7472)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7540..7558
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7596..7618)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7620..7640)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7625..7955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7806..7941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7868..7887
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8092..8111
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8178..8633)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8315..8336)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8527..8546
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8759..8781
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8773..8790
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8774..8790
                     /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 tgaatgttac
     1621 taaagatgag aacccaagat ccagaagtca agatcttcac ctttttcatg cttggatgat
     1681 gttaatcatg acggtgctct ttcttcctgt cactgaaacg tctaaacaaa atattccaag
     1741 actcaagcta acctacaaag acttgctgct ttcaaacacc tgtatcccct ttttgggttc
     1801 atcagaagga ttggatttcc agactcttct tttggatgag gagaggggca tactgctcct
     1861 aggagccaaa gaccatgtct tcctgctcag tctggttgac ttgaacaaga attttaagaa
     1921 gatttattgg cctgctgcaa aagaacgagt ggagctatgt aaattagctg ggaaagatgc
     1981 caatgcagaa tgtgcaaatt tcatccgtgt gcttcaaccc tataataaga ctcacgttta
     2041 cgtgtgtgga actggagcgt ttcatccgct gtgtgggtac attgatctcg gcgccaacaa
     2101 ggaggaactc atatttaaac tagacacgca caacctggag tctggcagac tgaaatgtcc
     2161 ctttgatcct cagcagcctt ttgcttcagt aatgacagat gagcacctct actctggaac
     2221 agcttctgat ttccttggca aagacactgc attcacaagg tctctggggc taatgcagga
     2281 ccaccattcc atcagaactg acatttcaga gcaccactgg ctcaatggag caaaatttat
     2341 cggaacattc cccattccag acacctataa tccagatgat gataaaatat atttcttctt
     2401 tcgagaatca tcccaggaag gcagtacttc tgacagaagc attctttcaa gagttggaag
     2461 agtttgtaag aatgatgtag gtgggcaacg aagtctgata aacaaatgga caacttttct
     2521 aaaggcaaga ctgatttgct cgattcctgg aagcgatggg gcagataccc attttgatga
     2581 actccaagat atttacttac tccctacgag agatgaaaga aatcctgtag tatatggagt
     2641 ctttaccaca accagctcca tcttcaaagg ctctgctgtc tgtgtgtaca gcatggctga
     2701 tatccgagca gtctttaatg gtccctatgc tcataaggaa agtgctgacc atcgctgggt
     2761 gcaatatgat ggaaggatac cttacccccg acctggaacg tgtccaagca aaacctatga
     2821 cccactgatt aagtccaccc gagactttcc agacgatgtt atcagtttca taaggcggca
     2881 ccctgtgatg tataagtccg tgtacccagt ggcaggagca ccgaccttca agagaatcaa
     2941 cgtggattac agactgacgc agatagtggt ggatcacgtg gtcgctgaag acgggcagta
     3001 tgatgtcatg tttctcggaa cagacattgg aacagtcctg aaagttgtga gcatctccaa
     3061 ggagaagtgg aatatggaag aggtcgtact ggaggagctt caggtattca agcacccaac
     3121 agctatcttg aacatggagt tgtcgctgaa gcagcaacag ttgtacgttg gttcctggga
     3181 tggattggtc cagctctcct tgcacagatg cgacacttac gggaaagcat gtgcagactg
     3241 ctgtctcgcc agagaccctt actgtgcctg ggatggaaat gcttgctcca gatatgcacc
     3301 cacgtctaaa aggcgagcta gacgccagga tgtaaaatat ggggacccaa tcactcagtg
     3361 ctgggacata gaagacagca ttagtcatga aacagctgat gaaaaggtga tttttggaat
     3421 tgaatttaat tcaacctttt tggagtgtat acctaaatcc caacaagcct ctgttgagtg
     3481 gtacatccag cggtcaggag atgagcatcg agaggagttg aaacctgatg aaaggatcat
     3541 caaaactgac tatgggctac tgattcgaag tctgcagaag aaggattctg ggatgtatta
     3601 ctgcaaagca caggagcaca ctttcatcca caccatagtg aagctgactt tgaatgtcat
     3661 tgagaatgaa cagatggaaa atacccagag ggcagaatac caggaggggc aggtcaagga
     3721 tctgttggct gagtcacggt tgagatacaa agactacatc caaatcctta gcagcccgaa
     3781 cttcagcctg gaccagtact gtgagcagat gtggtacaag gagaagcgga gacagcgcaa
     3841 caagggcagc ccaaagtgga agcacatgca ggaaatgaag aagaaaggcg cgcctggatc
     3901 ttccggaatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     3961 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     4021 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     4081 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     4141 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     4201 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     4261 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     4321 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     4381 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     4441 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     4501 gatcctaggt gggggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     4561 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     4621 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     4681 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     4741 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     4801 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     4861 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     4921 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     4981 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     5041 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     5101 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     5161 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     5221 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     5281 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     5341 ccccgccggc accaccgacg ccgcgcaccc gggtcatcat caccatcacc attaatctag
     5401 agatcttatt aaagcagaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5461 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5521 aactcatcaa tgtatcttat catgtctggt cgactctagc tagactcttc cgcttcctcg
     5581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7501 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7561 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7621 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     7681 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     7741 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     7801 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     7861 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     7921 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     7981 ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8041 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8101 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8161 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     8221 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     8281 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     8341 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     8401 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     8461 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     8521 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     8581 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     8641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     8701 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     8761 cagtcacgac gttgtaaaac gacggccagt gaatt
//
