LOCUS       sequence-138962-        8003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3D protein (truncated at cleavage site P3; ie, 
            long), 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 8003)
  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 8003)
  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: Sema3d(L)-Fc-His
FEATURES             Location/Qualifiers
     source          1..8003
                     /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             4583..4600
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4626..4747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4663..4682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4717..4736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4785..4802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4956..5544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5036..5055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5715..6506,6507..6575))
                     /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     6338..6357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6576..6680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6748..6766
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6804..6826)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6828..6848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6833..7163
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7014..7149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7076..7095
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7300..7319
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7386..7841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7523..7544)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7735..7754
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7967..7989
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7981..7998
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7982..7998
                     /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 tgaatgttac
     1621 taaagatgag aacccaagat ccagaagtca agatcttcac ctttttcatg cttggatgat
     1681 gttaatcatg acggtgctct ttcttcctgt cactgaaacg tctaaacaaa atattccaag
     1741 actcaagcta acctacaaag acttgctgct ttcaaacacc tgtatcccct ttttgggttc
     1801 atcagaagga ttggatttcc agactcttct tttggatgag gagaggggca tactgctcct
     1861 aggagccaaa gaccatgtct tcctgctcag tctggttgac ttgaacaaga attttaagaa
     1921 gatttattgg cctgctgcaa aagaacgagt ggagctatgt aaattagctg ggaaagatgc
     1981 caatgcagaa tgtgcaaatt tcatccgtgt gcttcaaccc tataataaga ctcacgttta
     2041 cgtgtgtgga actggagcgt ttcatccgct gtgtgggtac attgatctcg gcgccaacaa
     2101 ggaggaactc atatttaaac tagacacgca caacctggag tctggcagac tgaaatgtcc
     2161 ctttgatcct cagcagcctt ttgcttcagt aatgacagat gagcacctct actctggaac
     2221 agcttctgat ttccttggca aagacactgc attcacaagg tctctggggc taatgcagga
     2281 ccaccattcc atcagaactg acatttcaga gcaccactgg ctcaatggag caaaatttat
     2341 cggaacattc cccattccag acacctataa tccagatgat gataaaatat atttcttctt
     2401 tcgagaatca tcccaggaag gcagtacttc tgacagaagc attctttcaa gagttggaag
     2461 agtttgtaag aatgatgtag gtgggcaacg aagtctgata aacaaatgga caacttttct
     2521 aaaggcaaga ctgatttgct cgattcctgg aagcgatggg gcagataccc attttgatga
     2581 actccaagat atttacttac tccctacgag agatgaaaga aatcctgtag tatatggagt
     2641 ctttaccaca accagctcca tcttcaaagg ctctgctgtc tgtgtgtaca gcatggctga
     2701 tatccgagca gtctttaatg gtccctatgc tcataaggaa agtgctgacc atcgctgggt
     2761 gcaatatgat ggaaggatac cttacccccg acctggaacg tgtccaagca aaacctatga
     2821 cccactgatt aagtccaccc gagactttcc agacgatgtt atcagtttca taaggcggca
     2881 ccctgtgatg tataagtccg tgtacccagt ggcaggagca ccgaccttca agagaatcaa
     2941 cgtggattac agactgacgc agatagtggt ggatcacgtg gtcgctgaag acgggcagta
     3001 tgatgtcatg tttctcggaa cagacattgg aacagtcctg aaagttgtga gcatctccaa
     3061 ggagaagtgg aatatggaag aggtcgtact ggaggagctt caggtattca agcacccaac
     3121 agctatcttg aacatggagt tgtcgctgaa gcagcaacag ttgtacgttg gttcctggga
     3181 tggattggtc cagctctcct tgcacagatg cgacacttac gggaaagcat gtgcagactg
     3241 ctgtctcgcc agagaccctt actgtgcctg ggatggaaat gcttgctcca gatatgcacc
     3301 cacgtctaaa aggcgagcta gacgccagga tgtaaaatat ggggacccaa tcactcagtg
     3361 ctgggacata gaagacagca ttagtcatga aacagctgat gaaaaggtga tttttggaat
     3421 tgaatttaat tcaacctttt tggagtgtat acctaaatcc caacaagcct ctgttgagtg
     3481 gtacatccag cggtcaggag atgagcatcg agaggagttg aaacctgatg aaaggatcat
     3541 caaaactgac tatgggctac tgattcgaag tctgcagaag aaggattctg ggatgtatta
     3601 ctgcaaagca caggagcaca ctttcatcca caccatagtg aagctgactt tgaatgtcat
     3661 tgagaatgaa cagatggaaa atacccagag ggcagaatac caggaggggc aggtcaagga
     3721 tctgttggct gagtcacggt tgagatacaa agactacatc caaatcctta gcagcccgaa
     3781 cttcagcctg gaccagtact gtgagcagat gtggtacaag gagaagcgga gacagcgcaa
     3841 caagggcagc ccaaagtgga agcacatgca ggaaatgaag aagaaaggcg cgcctgacaa
     3901 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct
     3961 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
     4021 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
     4081 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
     4141 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
     4201 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
     4261 gccccgagaa ccacaggtgt ataccctgcc cccatcccgg gatgagctga ccaagaacca
     4321 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
     4381 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
     4441 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
     4501 cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
     4561 cctgtctccg ggtaaaggcg gccatcatca ccatcaccat taatctagag atcttattaa
     4621 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4681 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4741 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg
     4801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5641 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5701 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5761 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5821 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5881 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5941 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6001 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6061 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6121 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6181 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6241 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6301 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6361 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6421 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6481 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6541 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6601 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6661 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6721 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6781 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca
     6841 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     6901 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6961 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     7021 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     7081 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     7141 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg
     7201 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7261 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7321 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7381 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     7441 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     7501 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     7561 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7621 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7681 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7741 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7801 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     7861 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7921 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7981 tgtaaaacga cggccagtga att
//
