LOCUS       sequence-138624-        7763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4A 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 7763)
  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 7763)
  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: Sema4a-Fc-His
FEATURES             Location/Qualifiers
     source          1..7763
                     /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             4343..4360
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4386..4507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4423..4442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4477..4496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4545..4562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4716..5304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4796..4815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5475..6266,6267..6335))
                     /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     6098..6117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6336..6440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6508..6526
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6564..6586)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6588..6608)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6593..6923
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6774..6909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6836..6855
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7060..7079
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7146..7601)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7283..7304)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7495..7514
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7727..7749
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7741..7758
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7742..7758
                     /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 tggccctacc
     1621 atccctgggc caggactcat ggagtctcct gcgtgttttt ttcttccaac tcttcctgct
     1681 gccatcactg ccacctgctt ctgggactgg tggtcagggg cccatgccca gagtcaaata
     1741 ccatgctgga gacgggcaca gggccctcag cttcttccaa caaaaaggcc tccgagactt
     1801 tgacacgctg ctcctgagtg acgatggcaa cactctctat gtgggggctc gagaggccgt
     1861 cctggccttg aatatccaga acccaggaat cccaaggcta aagaacatga taccctggcc
     1921 agccagtgag agaaaaaaga ccgaatgtgc ctttaagaag aagagcaatg agacacagtg
     1981 tttcaacttc attcgagtcc tggtctctta caatgctact cacctctatg cctgtgggac
     2041 ctttgccttc agccctgcct gtaccttcat tgaactccaa gattccctcc tgttgcccat
     2101 cttgatagac aaggtcatgg acgggaaggg ccaaagcccc tttgaccctg ttcacaagca
     2161 cacagctgtc ttggtcgatg ggatgcttta ttccggcacc atgaacaact tcctgggcag
     2221 cgagcccatc ctgatgcgga cactgggatc ccagcctgtt ctcaagactg acatcttctt
     2281 acgctggctg cacgcggatg cctccttcgt ggcagccatt ccatccaccc aggtcgtcta
     2341 tttcttcttt gaggagacag ccagcgagtt tgacttcttt gaagagctgt atatatccag
     2401 ggtggctcaa gtctgcaaga acgacgtggg cggtgaaaag ctgctgcaga agaagtggac
     2461 caccttcctc aaagcccagt tgctctgcgc tcagccaggg cagctgccat tcaacatcat
     2521 ccgccacgcg gtcctgctgc ccgccgattc tccctctgtt tcccgcatct acgcagtctt
     2581 tacctcccag tggcaggttg gcgggaccag gagctcagca gtctgtgcct tctctctcac
     2641 ggacattgag cgagtcttta aagggaagta caaggagctg aacaaggaga cctcccgctg
     2701 gaccacttac cggggctcag aggtcagccc gaggccaggc agttgctcca tgggcccctc
     2761 ctctgacaaa gccttgacct tcatgaagga ccattttctg atggatgagc acgtggtagg
     2821 aacacccctg ctggtgaagt ctggtgtgga gtacacacgg cttgctgtgg agtcagctcg
     2881 gggccttgat gggagcagcc atgtggtcat gtatctgggt acctccacgg ggtccctgca
     2941 caaggctgtg gtgcctcagg acagcagtgc ttatctcgtg gaggagattc agctgagccc
     3001 tgactctgag cctgttcgaa acctgcagct ggcccccgcc cagggtgcag tgtttgcagg
     3061 cttctctgga ggcatctgga gagttcccag ggccaattgc agtgtctacg agagctgtgt
     3121 ggactgtgtg cttgccaggg accctcactg tgcctgggac cctgaatcaa gactctgcag
     3181 ccttctgtct ggctctacca agccttggaa gcaggacatg gaacgcggca acccggagtg
     3241 ggtatgcacc cgtggcccca tggccaggag cccccggcgt cagagccccc ctcaactaat
     3301 taaagaagtc ctgacagtcc ccaactccat cctggagctg ccctgccccc acctgtcagc
     3361 actggcctct taccactgga gtcatggcca agccaaaatc tcagaagcct ctgctaccgt
     3421 ctacaatggc tccctcttgc tgctgccgca ggatggtgtc gggggcctct accagtgtgt
     3481 ggcgactgag aacggctact cataccctgt ggtctcctat tgggtagaca gccaggacca
     3541 gcccctggcg ctggaccctg agctggcggg cgttccccgt gagcgtgtgc aggtcccgct
     3601 gaccagggtc ggaggcggag cttccatggc tgcccagcgg tcctacggcg cgcctgacaa
     3661 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct
     3721 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
     3781 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
     3841 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
     3901 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
     3961 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
     4021 gccccgagaa ccacaggtgt ataccctgcc cccatcccgg gatgagctga ccaagaacca
     4081 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
     4141 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
     4201 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
     4261 cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
     4321 cctgtctccg ggtaaaggcg gccatcatca ccatcaccat taatctagag atcttattaa
     4381 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4441 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4501 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg
     4561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6481 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6541 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca
     6601 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     6661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6841 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6901 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg
     6961 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7021 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7081 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7141 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     7201 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     7261 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     7321 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7381 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7441 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7501 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7561 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     7621 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7681 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7741 tgtaaaacga cggccagtga att
//
