LOCUS       sequence-138687-        7375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4F, isoform b 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 7375)
  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 7375)
  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: Sema4f.b-Fc-His
FEATURES             Location/Qualifiers
     source          1..7375
                     /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             3955..3972
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3998..4119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4035..4054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4089..4108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4157..4174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4328..4916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4408..4427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5087..5878,5879..5947))
                     /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     5710..5729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5948..6052)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6120..6138
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6176..6198)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6200..6220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6205..6535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6386..6521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6448..6467
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6672..6691
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6758..7213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6895..6916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7107..7126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7339..7361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7353..7370
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7354..7370
                     /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 gccaccatgc tggccagggc cgagcggccc
     1621 cgcccgggcc cccggccgcc tccggtctct ctcttcccgc cgccgtcgtc gctgctgctg
     1681 ctgctgctgg cgatgctgag cgccccggtg tgcggccgcg tcccccgctc ggtgcccaga
     1741 acctcgctgc ccatctccga ggctgactcc tatctcaccc ggtttgcggc ccctcatacg
     1801 tacaattact ctgctctcct ggtggatcct gcttctcaca cactttacgt cggtgcacgg
     1861 gacagcatct tcgctttaac cctgcccttc tctggggaaa agcctcgaag gatcgactgg
     1921 atggtgcccg agactcacag acagaactgt aggaagaaag gcaagaaaga ggacgaatgt
     1981 cacaatttta tccagattct cgccattgcc aatgcctctc acctcctcac gtgtggcacc
     2041 ttcgcttttg atccgaagtg cggggttatt gatgtgtcca gtttccagca ggttgaaaga
     2101 cttgagagtg gccgggggaa atgtcctttt gagccagctc aacggtcagc agctgtaatg
     2161 gctgggggcg tcctctacac cgccactgtg aaaaacttcc tggggacaga gccgattatc
     2221 tcccgagctg tgggtcgagc tgaggactgg attcgaacag agaccttgtc atcctggctt
     2281 aatgccccag cctttgtcgc agctatggtc ctgagcccgg ctgagtgggg ggatgaagat
     2341 ggagacgatg aaatcttctt tttcttcacg gagacctccc gagtcttgga ctcatatgag
     2401 cgcatcaagg tcccacgagt ggcccgtgtg tgtgcggggg accttggggg ccggaagacc
     2461 cttcagcaga gatggacgac gtttctaaag gctgacctgc tgtgtccagg gcccgagcat
     2521 ggaagggcct cgggggttct gcaggatatg acagagcttc gacctcagcc tggcgcgggg
     2581 acccccctct tttatggcat cttttcctcc cagtgggaag gagccgccat ctctgctgtg
     2641 tgtgccttcc gaccccaaga catccgggca gtgctgaatg gtccctttag agagctaaaa
     2701 catgactgca acaggggact gcctgtcatg gacaacgagg tgccccagcc cagacctgga
     2761 gagtgcatca ccaacaacat gaagttccag cagtttggat cctcactctc cctgccagac
     2821 cgtgtgctca cctttatcag agaccacccg ctcatggaca ggcccgtgtt tccagctgat
     2881 ggccgccccc tgctggtcac tacggataca gcctatctca gagttgtggc tcaccgggtg
     2941 accagcctct cagggaaaga atatgatgtg ctctacctag ggacagagga tgggcacctc
     3001 caccgggctg tgcgcatcgg agctcagctc agtgtcctgg aggatctggc cttgttcccg
     3061 gaaccacaac cggttgagag catgaaattg taccatgatt ggctcctggt gggctcccat
     3121 actgaggtga cacaagtgaa caccagcaac tgtggccgtc tccagagctg ctcagaatgt
     3181 atcctggccc aggatcccgt gtgtgcctgg agcttccggc tcgatgcttg tgtggcccat
     3241 gcaggcgagc accgtggggg cgcgcctgac aaaactcaca catgcccacc gtgcccagca
     3301 cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa ggacaccctc
     3361 atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct
     3421 gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa gacaaagccg
     3481 cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag
     3541 gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct cccagccccc
     3601 atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt gtataccctg
     3661 cccccatccc gggatgagct gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc
     3721 ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga gaacaactac
     3781 aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag caagctcacc
     3841 gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat gcatgaggct
     3901 ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaagg cggccatcat
     3961 caccatcacc attaatctag agatcttatt aaagcagaac ttgtttattg cagcttataa
     4021 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4081 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggt cgactctagc
     4141 tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5701 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5761 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5821 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5881 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5941 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6001 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6061 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6121 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6181 gacacatgca gctcccggag gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     6241 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     6301 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6361 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6421 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     6481 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6541 cccgggagct tgtatatcca ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga
     6601 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     6661 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     6721 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     6781 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6841 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6901 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6961 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7021 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7081 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7141 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7201 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     7261 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     7321 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaatt
//
