LOCUS       sequence-138975-        8636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema5A protein, 
            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 8636)
  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 8636)
  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: Sema5a-Fc-His
FEATURES             Location/Qualifiers
     source          1..8636
                     /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             5216..5233
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5259..5380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5296..5315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5350..5369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5418..5435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5589..6177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5669..5688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6348..7139,7140..7208))
                     /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     6971..6990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7209..7313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7381..7399
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7437..7459)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7461..7481)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7466..7796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7647..7782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7709..7728
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7933..7952
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8019..8474)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8156..8177)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8368..8387
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8600..8622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8614..8631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8615..8631
                     /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 tgaagggagc
     1621 ctgcatcctt gcatggctgt tctcaagcct gggagtgtgg agacttgcta ggcccgagac
     1681 ccaggaccct gccaagtgcc agagagctga gcaccctgtc gtctcttaca aagaaattgg
     1741 cccctggtta cgggagttca gagccgagaa tgctgtggat ttctcgaggt taacatttga
     1801 cccaggacag aaagaacttg tcgtaggagc gagaaactat ctcttcagat tagagcttga
     1861 ggatctgtct ctcatccagg ctgtggagtg ggagtgtgac gaagccacca agaaggcctg
     1921 ttacagcaag ggcaaatcga aggaggaatg tcagaactac atacgagtgc tcttagttgg
     1981 tggggaccgg ctattcacct gtgggaccaa tgccttcaca cctgtctgta ccatccgctc
     2041 gttaagtaac ctgactgaga tccatgatca gatcagtggc atggcccgct gtccctacag
     2101 tccccagcac aattctaccg ccctactcac tgccagtggt gagctctatg ctgcaacagc
     2161 catggatttc ccagggcgag atccagccat ttaccgaagt ctgggcactt tgcctcctct
     2221 tcgaactgca cagtacaact ccaaatggct caatgaacca aactttgtgt cttcctacga
     2281 catcggaaat ttcacctact tcttcttccg agaaaacgcc gtggaacatg actgtgggaa
     2341 gacggtgttc tccagggctg cccgggtctg taaaaatgac attggagggc gatttcttct
     2401 ggaggacacc tggactacct tcatgaaggc tcgcctcaac tgctcccggc ctggtgaggt
     2461 gcccttctac tacaatgagc tgcagggtac tttcttcctg ccggagctgg atctgatcta
     2521 tggtatcttc accaccaatg taaacagcat cgctgcctct gctgtctgtg tcttcaacct
     2581 gagcgccatc tcacaggcct tcaatggacc cttcaagtac caagaaaact ctcgctcagc
     2641 ctggctgcct tatcccaacc ccaaccccaa ttttcagtgc ggcaccatgg accagggcct
     2701 gtacgtaaac ctgacggaaa gaaacctaca ggatgctcag aagttcattc tgatgcatga
     2761 agtggtgcag ccagtgacca cagtgccctc tttcatggag gacaacagcc gcttctccca
     2821 cttagctgtt gatgttgtgc aaggcaggga gactctggtc cacatcattt atctggccac
     2881 agattatggc accattaaga aagtacgagc tcccctgagt cagagctcag gcagctgttt
     2941 gctggaagag attgagcttt tcccagagag gaggagtgag cccatcagga gcctgcagat
     3001 cctccacagc cagagtgtcc tatttgtggg actacaggag catgtggcca agatccccct
     3061 gaagaggtgc cacttccatc aaacacgcag tgcctgcatt ggcgctcagg acccttactg
     3121 tggctgggat gcggtgatga agaaatgcac cagcctggag gagagtctga gcatgaccca
     3181 gtgggatcag agcatcccca cctgtccgac cagaaatctc actgtagatg ggagctttgg
     3241 cccatggtca ccgtggacac cctgtacaca cactgatggc actgctgtgg gctcctgcct
     3301 ctgccggtcc cgctcctgtg acagcccagc tcctcagtgt ggtggttggc agtgtgaggg
     3361 ccctagaatg gagatcacca actgttccag gaatggaggc tggactccct ggacctcctg
     3421 gtctccgtgc agcacgacct gtggcattgg cttccaagtg cggcagcgat cctgcagcaa
     3481 ccccacgccc aggcatggcg ggcgcgtgtg cgtgggccag aacagggagg aaagatactg
     3541 caatgaacat ttgctctgcc caccacacgt gttctggaca ggctggggac cttgggaacg
     3601 gtgcacagcc cagtgcgggg gtggcattca agcccgccgg aggacctgtg aaaatgggcc
     3661 tgactgtgca ggatgcaatg tggaatacca gccttgtaac accaatgcat gcccggagct
     3721 gaagaagacc acaccctgga caccctggac ccctgtcaac atctctgaca atggaggtca
     3781 ctatgagcag cgtttccgct atacctgtaa agctcgcctg ccagatccaa atttgctgga
     3841 agtaggaaga cagaggatag aaatgcggta ctgttccagc gatggaacca gtggctgctc
     3901 cacagacgga ctttctggag actttctaag agctgggaga tactctgctc atacagtcaa
     3961 tggggcatgg tcagcctgga cttcctggtc acagtgcagc agagactgca gcaggggcat
     4021 tcggaaccgg aagcgtgttt gcaacaaccc agaacccaag tttgggggca tgccatgcct
     4081 tggcccatcg ctggagttcc aggaatgcaa cattttacct tgtccagtgg atggtgtgtg
     4141 gtcttgctgg tcatcctggt ctaaatgttc agcaacctgt ggaggtggcc actacatgag
     4201 aacccgttct tgttcgaatc cagccccagc atatggaggg gacatctgcc tgggactgca
     4261 cacagaagag gcactctgca acacacagac ctgcccagaa agctggtcag agtggtcaga
     4321 ctggtctgtg tgcgatgcat ctggtaccca ggtccgtgct cggcagtgca tccttctgtt
     4381 tccagtgggc agccaatgtt ctggaaatac tacagagagc cggccttgtg tatttgactc
     4441 taatttcatc ccagaagtat ctgtggcaag atccagcagc gtagaagaaa aaaggtgtgg
     4501 agagttcaac atgttccacg gcgcgcctga caaaactcac acatgcccac cgtgcccagc
     4561 acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct
     4621 catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc
     4681 tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc
     4741 gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca
     4801 ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc tcccagcccc
     4861 catcgagaaa accatctcca aagccaaagg gcagccccga gaaccacagg tgtataccct
     4921 gcccccatcc cgggatgagc tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg
     4981 cttctatccc agcgacatcg ccgtggagtg ggagagcaat gggcagccgg agaacaacta
     5041 caagaccacg cctcccgtgc tggactccga cggctccttc ttcctctaca gcaagctcac
     5101 cgtggacaag agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggc
     5161 tctgcacaac cactacacgc agaagagcct ctccctgtct ccgggtaaag gcggccatca
     5221 tcaccatcac cattaatcta gagatcttat taaagcagaa cttgtttatt gcagcttata
     5281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg tcgactctag
     5401 ctagactctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5461 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5521 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5581 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5641 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5701 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5761 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5821 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5881 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5941 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6001 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6061 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6481 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7321 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     7381 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     7441 tgacacatgc agctcccgga ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     7501 cccaggcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     7561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     7621 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     7681 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     7741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     7801 tcccgggagc ttgtatatcc attttcgtcc cggagacggt cacagcttgt ctgtaagcgg
     7861 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     7921 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     7981 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     8041 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     8101 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     8161 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     8221 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     8281 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     8341 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     8401 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     8461 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     8521 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     8581 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaatt
//
