LOCUS       sequence-138677-        7826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4B 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 7826)
  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 7826)
  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: Sema4b-Fc-His
FEATURES             Location/Qualifiers
     source          1..7826
                     /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             complement(2807..2818)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             4406..4423
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4449..4570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4486..4505)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4540..4559
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4608..4625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4779..5367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4859..4878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5538..6329,6330..6398))
                     /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     6161..6180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6399..6503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6571..6589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6627..6649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6651..6671)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6656..6986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6837..6972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6899..6918
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7123..7142
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7209..7664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7346..7367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7558..7577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7790..7812
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7804..7821
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7805..7821
                     /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 tgggccgggc
     1621 gagccgctcc gctgtcctgc ggcgcgcgct gctgctgctg ctgctactgc tgctgctgcg
     1681 gacgacgact acgcgggcgc tcggcccccg gatcagtgtg ccgctgggct ctgaagaaag
     1741 gctgattaga aaatttgaag ctgaaaacat ctccaactac acggcccttc tgctgagcca
     1801 ggatggcaag acgctgtatg tgggggcccg agaggccctc tttgcactta acagcaacct
     1861 cagcttcttg ccaggcgggg agtaccaaga gctgctgtgg agtgcagatg ctgacaggaa
     1921 gcagcagtgc agcttcaagg gcaaggaccc aaagcgtgac tgtcaaaact acatcaagat
     1981 cctcctgcca ctcaacagca gccacctgct cacctgtggc acggccgcct tcagccccct
     2041 gtgtgcttac attcacatag cgagctttac tttagcccaa gatgaggccg gcaatgtcat
     2101 tctggaggat ggcaagggtc gttgtccctt tgaccccaac ttcaagtcca cggctctggt
     2161 ggttgatggt gagctgtaca ctggaacagt cagtagcttc cagggaaacg acccagccat
     2221 ttcccggagc cagagttccc gccccaccaa gactgagagc tccctcaact ggctacaaga
     2281 ccctgccttt gtggcctcgg cctacgtccc cgagagcctg ggcagcccca taggtgatga
     2341 tgataagatc tacttcttct tcagcgagac gggccaggag tttgagttct ttgagaacac
     2401 catcgtgtcc cgagttgccc gagtctgtaa gggcgatgag ggtggagagc gggtgttgca
     2461 gcaacgctgg acctcctttc tcaaggctca gctcctgtgc tcccggcctg atgatggctt
     2521 tccctttaac gtgctacaag atgtcttcac cctgaacccc aaccctcagg attggcgcaa
     2581 gacccttttc tatggggtct ttacctccca gtggcacaga gggaccacag aaggctctgc
     2641 catctgcgtc ttcaccatga atgatgtgca gaaggccttt gacggcctgt acaagaaagt
     2701 aaacagagag acacagcagt ggtataccga gacccaccag gtgcccacac cgcggccggg
     2761 agcgtgcatt accaacagtg cccgggaacg gaagatcaac tcgtccctgc agctcccaga
     2821 ccgagtgctg aacttcctca aggatcactt cttgatggat gggcaggtcc gcagtcgcct
     2881 gctgctgctg cagcccagag cccgctacca gcgtgtggct gtgcaccgtg tgcctggcct
     2941 gcacagcact tatgatgtcc tatttctggg cactggtgat ggccgcctgc acaaagcagt
     3001 gaccctgagc tccagagtcc acatcattga ggagctgcag atcttccctc aaggacagcc
     3061 tgtgcagaac ctgctcttgg acagccatgg gggactgttg tatgcctcct cccattccgg
     3121 ggtggtgcaa gtgcccgtag ccaactgcag cctgtaccca acctgtggag actgcctcct
     3181 ggctcgagac ccctactgcg cctggactgg ctctgcctgc aggctcgcta gcctctacca
     3241 gcctgatctg gcctccaggc catggaccca ggacattgag ggtgccagtg tcaaggaact
     3301 ctgcaagaat tcctcataca aggcccggtt tcttgtgcca ggtaagccat gtaaacaagt
     3361 ccagatccaa ccaaacacag tgaacaccct ggcctgccca ctcctctcaa acctggccac
     3421 tcggctctgg gtgcacaatg gagccccagt caatgcctct gcctcctgcc gcgtgttacc
     3481 caccggggac ctgctgctgg tgggcagcca gcagggtttg ggggtgttcc agtgttggtc
     3541 gatagaagaa ggattccagc agcttgtggc cagctactgc ccagaggtga tggaggaggg
     3601 ggtaatggac caaaagaacc agcgtgatgg taccccagtc attatcaaca catcacgagt
     3661 gagtgcaccg gctggtggca gggccagctg gggtgcggac aagtcctacg gcgcgcctga
     3721 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     3781 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     3841 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     3901 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     3961 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     4021 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     4081 gcagccccga gaaccacagg tgtataccct gcccccatcc cgggatgagc tgaccaagaa
     4141 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     4201 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     4261 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     4321 cgtcttctca tgctccgtga tgcatgaggc tctgcacaac cactacacgc agaagagcct
     4381 ctccctgtct ccgggtaaag gcggccatca tcaccatcac cattaatcta gagatcttat
     4441 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4501 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4561 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac
     4621 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4681 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4741 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4801 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4861 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4921 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4981 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5041 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5101 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5161 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5221 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5281 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5341 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5401 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5461 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5521 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5581 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5641 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5701 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5761 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5821 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5881 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5941 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6001 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6061 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6121 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6181 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6241 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6301 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6361 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6421 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6481 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6541 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6601 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg
     6661 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     6721 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     6781 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6841 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6901 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     6961 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc
     7021 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     7081 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     7141 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7201 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7261 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     7321 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     7381 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     7441 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     7501 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     7561 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     7621 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     7681 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     7741 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     7801 cgttgtaaaa cgacggccag tgaatt
//
