LOCUS       sequence-138692-        8654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema5B 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 8654)
  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 8654)
  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: Sema5b-Fc-His
FEATURES             Location/Qualifiers
     source          1..8654
                     /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             5234..5251
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5277..5398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5314..5333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5368..5387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5436..5453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5607..6195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5687..5706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6366..7157,7158..7226))
                     /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     6989..7008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7227..7331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7399..7417
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7455..7477)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7479..7499)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7484..7814
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7665..7800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7727..7746
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7951..7970
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8037..8492)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8174..8195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8386..8405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8618..8640
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8632..8649
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8633..8649
                     /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 tggtagtccc
     1621 aggacccctg gctctctcgc tgttgctgtc cagcctcacc ctgctggtgt cccacctctc
     1681 cagctcccag gacattgcca gtgagtccag cagtgagcaa cagatgtgca cgcggaggga
     1741 gcaccccatc gtggcctttg aagatctgaa gccgtgggtc tttaacttca cctaccctgg
     1801 agtccgggat ttctcccagc ttgctcttga tccctcgagg aatcagctca tcgtgggagc
     1861 caggaactac ctcttcagac tcagcctcgc caatgtctcc ctccttcagg ctacagagtg
     1921 ggcctccagt gaggacacgc gccgctcctg ccagagcaaa gggaaaacgg aggaggagtg
     1981 tcagaactat gtacgagtcc tgattgtttc cggccggaag gtgttcatgt gcggtaccaa
     2041 tgccttttcc ccagtgtgct ccagcagaca ggtagggaac ctcagccgga ctattgagaa
     2101 gatcaacggt gtggcccggt gcccctatga cccacgccac aactctacag ccgtcatctc
     2161 ctctcagggg gagctctatg cagccacagt cattgacttc tccggtcggg acccagccat
     2221 ctaccgcagc ctgggcagtg ggccaccgct tcgcactgcc cagtataact ccaaatggct
     2281 caatgagcca aattttgtgg cagcctttga catcgggctg tttgcgtatt tcttccttcg
     2341 ggagaatgcc gtggagcatg attgtgggcg cactgtgtac tctcgggtgg cccgggtgtg
     2401 caagaatgat gtaggtggcc gtttcctgct ggaggacacg tggaccacat tcatgaaggc
     2461 ccggctcaac tgctcccgcc cgggagaggt ccccttctac tataatgagc tgcagagtgc
     2521 cttccatctg cccgagcagg acctcatcta tggcgtcttc accactaacg taaacagcat
     2581 tgcggcttct gctgtctgcg ccttcaacct cagtgccatc tccaaggctt tcaatggccc
     2641 atttcgttac caggaaaacc ccagggctgc ctggctccca atcgccaatc ccatccccaa
     2701 tttccagtgt ggcactctgc cggagactgg ccccaacgag aacctcacgg agcgcagcct
     2761 gcaggacgca cagcggctct tcctgatgag cgaagctgtg cagccagtga caccagagcc
     2821 ctgtgtcacc caggacagcg tccgcttctc acatctcgtg gtagaccttg tgcaagctaa
     2881 ggacacgctc tatcacgtac tctacatagg cacggagtcg ggtaccatcc tgaaagcgct
     2941 gtccacggcc agccgcagcc tccgtggctg ctacctggag gagctacatg tgctgcctcc
     3001 tgggcgcctt gaacctctgc ggagcctgcg catcctgcac agcgcgcgtg cgctcttcgt
     3061 gggattgagc gacagggtgc tgcgggtccc actggagagg tgttcggcct atcatagcca
     3121 gggggcatgc ctgggagcac gggacccata ctgcggctgg gatgggaagc ggcaactttg
     3181 cagcacgctt gaagacagtt ccaacatgag cctgtggatc cagaacatca caacctgtcc
     3241 tgtacgaaat gtgacacggg atgggggctt cggcccatgg tcaccatgga aaccgtgtga
     3301 gcacttagat ggagacaact cgggttcttg cctgtgccgg gccagatcct gtgactcccc
     3361 aaggcctcgc tgtgggggcc tcgagtgcct ggggccatcc atccatattg ccaattgctc
     3421 caggaatggg gcgtggaccg catggtcatc gtgggctcag tgcagcacgt cctgtgggat
     3481 cggcttccag gtccgtcagc gaagttgcag caacccggcg ccccgccatg ggggccgcat
     3541 ttgcgtgggc aagagccggg aggagcggtt ctgtaatgaa aatacacctt gcccagtgcc
     3601 catcttctgg gcttcctggg gttcctggag caagtgcagc aacaactgtg gaggcggcgt
     3661 gcagtcgcga cgccgttctt gcgagaatgg caactcgtgc ccgggttgcg gcgtggagtt
     3721 caagacctgc aaccccgagg cttgcccgga agtgcgacgc aacacaccct ggacgccctg
     3781 gctgcccgtg aacgtgaccc agggtggagc gcgccaggag cagcgattcc gcttcacctg
     3841 ccgcgcgccg ctgccggacc cgcacggtct gcagttcggc aagaggagga cagagaccag
     3901 gacttgcccg gcagacggca ccggagcctg cgacaccgac gccctggtgg aggatctcct
     3961 gcgcagcggg agcacgtcac cacacactct gaacggaggc tgggccacct ggggcccgtg
     4021 gtcatcctgc tcccgggact gcgagctggg cttccgcgtc cgcaagagaa cttgtaccaa
     4081 cccggagcct cgcaacgggg gcttgccctg cgtgggagac gctgcggagt accaagactg
     4141 caacccacag gcttgcccag tgcggggtgc ttggtcctgc tggaccgcat ggtcccagtg
     4201 ctcagcatcc tgcggtggtg gccactatca acgcacccgt tcctgcacca gccccgcccc
     4261 atccccaggt gaggacatct gcctcggcct gcacacggag gaggccctat gttcaacaca
     4321 ggcctgccca gaaggctggt cactgtggtc tgagtggggt gtctgcactg aggatggggc
     4381 ccagagccgg agccggagct gtgaggagct tctcccagga ccaggtgcct gtgttggcaa
     4441 cagcagccag agccggccct gcccctacag tgagattcct gtcatcctac ctgcttccag
     4501 tgtggaggag accaccagct gtggagggtt caatctcggc gcgcctgaca aaactcacac
     4561 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     4621 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     4681 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     4741 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     4801 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     4861 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     4921 accacaggtg tataccctgc ccccatcccg ggatgagctg accaagaacc aggtcagcct
     4981 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     5041 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     5101 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     5161 ctccgtgatg catgaggctc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     5221 gggtaaaggc ggccatcatc accatcacca ttaatctaga gatcttatta aagcagaact
     5281 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5341 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5401 atgtctggtc gactctagct agactcttcc gcttcctcgc tcactgactc gctgcgctcg
     5461 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5521 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5581 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5641 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5701 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5761 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5821 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5881 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5941 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6001 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6061 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6121 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6181 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6241 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6301 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6361 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6421 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6481 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6541 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6601 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6661 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6721 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6781 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6841 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6901 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6961 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7021 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7081 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7141 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7201 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7261 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7321 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     7381 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     7441 gatgacggtg aaaacctctg acacatgcag ctcccggagg aatgtgtgtc agttagggtg
     7501 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     7561 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     7621 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     7681 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7741 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7801 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcgtcccg gagacggtca
     7861 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     7921 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     7981 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     8041 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     8101 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     8161 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     8221 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     8281 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     8341 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     8401 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8461 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     8521 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     8581 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8641 acggccagtg aatt
//
