LOCUS       sequence-138947-        7922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4D 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 7922)
  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 7922)
  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: Sema4d-Fc-His
FEATURES             Location/Qualifiers
     source          1..7922
                     /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             4502..4519
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4545..4666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4582..4601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4636..4655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4704..4721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4875..5463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4955..4974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5634..6425,6426..6494))
                     /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     6257..6276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6495..6599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6667..6685
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6723..6745)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6747..6767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6752..7082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6933..7068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6995..7014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7219..7238
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7305..7760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7442..7463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7654..7673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7886..7908
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7900..7917
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7901..7917
                     /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 tgaggatgtg
     1621 tgcccccgtt agggggctgt tcttggccct ggtggtagtg ttgagaaccg cggtggcatt
     1681 tgcacctgtg cctcggctca cctgggaaca tggagaggta ggtctggtgc agtttcacaa
     1741 gccaggcatc tttaactact cggccttgct gatgagtgag gacaaagaca ctctgtatgt
     1801 aggcgcccgg gaagcagtct ttgcagtgaa tgcgctgaac atctctgaga agcaacatga
     1861 ggtatattgg aaggtctctg aagacaaaaa atccaagtgt gcagagaagg ggaaatcaaa
     1921 gcagacggaa tgcctaaact acattcgagt actacagcca ctaagcagca cttccctcta
     1981 tgtgtgtggg accaatgcgt tccagcccac ctgtgaccac ctgaacttga catccttcaa
     2041 gtttctgggg aaaagtgaag atggcaaagg aagatgcccc ttcgaccccg cccacagcta
     2101 cacatcagtc atggttgggg gcgagctcta ctctgggacg tcctataatt tcttgggcag
     2161 tgaacccatc atctctcgaa actcttccca cagtcccttg aggacggagt atgccatccc
     2221 gtggctgaac gagcctagct tcgtctttgc tgacgtgatc cagaaaagcc cagatggtcc
     2281 ggagggtgaa gatgacaagg tctacttctt ttttacggag gtatccgtgg agtacgaatt
     2341 cgtcttcaag ttgatgatcc cgcgagttgc cagggtgtgc aagggcgacc agggcggcct
     2401 gcggactttg caaaaaaagt ggacctcctt cctaaaggcc aggctgatct gctccaagcc
     2461 agacagtggc ctggtcttca acatacttca ggatgtgttt gtgctgaggg ccccgggcct
     2521 caaggagcct gtgttctatg cggtcttcac cccacagctg aacaatgtgg gtctgtcagc
     2581 ggtgtgcgcc tacacactgg ccacggtgga ggcagtcttc tcccgtggaa agtacatgca
     2641 gagtgccaca gtggagcagt ctcacaccaa gtgggtgcgc tacaatggcc cagtgcccac
     2701 tccccgacct ggagcgtgta tcgacagtga ggcccgggca gccaactaca ccagctcctt
     2761 gaatctccca gacaaaacac tgcagtttgt aaaagaccac cctttgatgg atgactcagt
     2821 gaccccgata gacaacagac ccaagctgat caaaaaagat gtaaactaca cccagatagt
     2881 ggtagacagg acccaggccc tggatgggac tttctacgac gtcatgttca tcagcacaga
     2941 ccggggagct ctgcataaag cagtcatcct cacaaaagag gtgcatgtca tcgaggagac
     3001 ccaactcttc cgggactctg aaccggtcct aactctgctg ctatcgtcaa agaaggggag
     3061 gaagtttgtc tatgcaggct ccaactctgg agtggtccaa gcgcccctgg cattctgcga
     3121 aaagcacggt agctgtgaag actgtgtgtt agcacgggac ccctactgtg cctggagccc
     3181 agccatcaag gcctgtgtta ccctgcacca ggaagaggcc tccagcaggg gctggattca
     3241 ggacatgagc ggtgacacat cctcatgcct ggataagagt aaagaaagtt tcaaccagca
     3301 ttttttcaag cacggcggca cagcggaact caaatgtttc caaaagtcca acctagcccg
     3361 ggtggtatgg aagttccaga atggcgagtt gaaggccgca agtcccaagt acggctttgt
     3421 gggcaggaag cacctgctca tcttcaacct gtcggacgga gacagcggcg tgtaccagtg
     3481 cctgtcagag gaaagggtga ggaataaaac ggtctcccag ctgctggcca agcacgttct
     3541 ggaagtgaag atggtacctc ggaccccccc ctcacctacc tcagaggatg ctcagacaga
     3601 aggtagtaag atcacatcca aaatgccggt tgcatctacc caggggtcct ctccccctac
     3661 cccggctctg tgggcaacct cccccagagc cgccacccta cctcccaagt cctcctccgg
     3721 cacatcctgt gaaccaaaga tggtcatcaa cacggtcccc cagctccact cagagaagac
     3781 ggtgtatctc aagtccagtg acaacggcgc gcctgacaaa actcacacat gcccaccgtg
     3841 cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga
     3901 caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga
     3961 agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac
     4021 aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct
     4081 gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc
     4141 agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta
     4201 taccctgccc ccatcccggg atgagctgac caagaaccag gtcagcctga cctgcctggt
     4261 caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa
     4321 caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa
     4381 gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca
     4441 tgaggctctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaaggcgg
     4501 ccatcatcac catcaccatt aatctagaga tcttattaaa gcagaacttg tttattgcag
     4561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggtcga
     4681 ctctagctag actcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4741 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4801 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4861 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4921 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4981 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5041 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5101 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5161 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5221 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5281 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5341 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5401 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5461 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5521 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5641 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5701 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5761 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5821 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5881 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5941 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6001 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6061 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6481 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6541 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6601 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6661 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6721 aacctctgac acatgcagct cccggaggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     6781 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     6841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7021 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     7081 aaaagctccc gggagcttgt atatccattt tcgtcccgga gacggtcaca gcttgtctgt
     7141 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     7201 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     7261 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     7321 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     7381 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     7441 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     7501 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     7561 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     7621 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     7681 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     7741 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7801 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     7861 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     7921 tt
//
