LOCUS       sequence-138971-        7373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3A protein (truncated at cleavage site P1; ie, 
            short), C-terminally fused to the Fc region of human IgG1 + 6X 
            histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7373)
  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 7373)
  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(S)-Fc-His
FEATURES             Location/Qualifiers
     source          1..7373
                     /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             3953..3970
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3996..4117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4033..4052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4087..4106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4155..4172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4326..4914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4406..4425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5085..5876,5877..5945))
                     /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     5708..5727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5946..6050)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6118..6136
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6174..6196)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6198..6218)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6203..6533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6384..6519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6446..6465
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6670..6689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6756..7211)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6893..6914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7105..7124
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7337..7359
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7351..7368
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7352..7368
                     /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 actgcaggcg cgcctgacaa aactcacaca tgcccaccgt gcccagcacc
     3301 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     3361 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     3421 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     3481 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     3541 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     3601 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt ataccctgcc
     3661 cccatcccgg gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     3721 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     3781 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     3841 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct
     3901 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaaggcg gccatcatca
     3961 ccatcaccat taatctagag atcttattaa agcagaactt gtttattgca gcttataatg
     4021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagcta
     4141 gactcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4201 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4261 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4321 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4381 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4441 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4501 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4561 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4621 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4681 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4741 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4801 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4861 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4921 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4981 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5041 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5101 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5161 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5221 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5281 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5341 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5401 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5461 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5521 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5581 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5641 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5701 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5761 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5821 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5881 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5941 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6001 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6061 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6121 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6181 cacatgcagc tcccggagga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6241 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6361 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     6481 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     6541 cgggagcttg tatatccatt ttcgtcccgg agacggtcac agcttgtctg taagcggatg
     6601 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     6661 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     6721 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     6781 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6841 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6901 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6961 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     7021 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     7081 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     7141 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     7201 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     7261 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     7321 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
