LOCUS       sequence-131206-        8180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3E protein (truncated at cleavage site P1; ie, 
            short), C-terminally fused to alkaline phosphatase + 6X histidine 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8180)
  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 8180)
  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: Sema3e(S)-AP-His
FEATURES             Location/Qualifiers
     source          1..8180
                     /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             4760..4777
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4803..4924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4840..4859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4894..4913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4962..4979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5133..5721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5213..5232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5892..6683,6684..6752))
                     /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     6515..6534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6753..6857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6925..6943
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6981..7003)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7005..7025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7010..7340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7191..7326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7253..7272
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7477..7496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7563..8018)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7700..7721)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7912..7931
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8144..8166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8158..8175
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8159..8175
                     /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 tggcaccggc
     1621 cggacacatc ctcaccttgc tgctctgggg tcacctgctg gaactctgga ccccaggtca
     1681 ctccgcgaac ccctcctacc ccaggctacg cctgtcacat aaagaacttt tggaactgaa
     1741 taggacttca atatttcaaa gcccccttgg atttcttgat ctccatacaa tgctgctgga
     1801 tgagtatcaa gaacggctct ttgtgggagg cagagacctt gtctattccc tgaacttgga
     1861 acgagtcagt gacggctaca gagagatata ctggccgagc acagcagtaa aggtagaaga
     1921 atgcataatg aaaggaaaag acgcaaatga gtgtgccaat tatatccggg ttttgcatca
     1981 ctacaacagg acacaccttc tgacctgtgc tactggagct tttgatccac actgtgcctt
     2041 catcagagtc gggcaccatt cagaggaacc cctgtttcac ctggagtcac acagatctga
     2101 gagaggaagg ggcagatgtc cttttgaccc caactcctcc tttgtgtcca cgctagttgg
     2161 gaatgagctg tttgctggac tctacagtga ctattggggc agagactcgg cgatcttccg
     2221 cagcatgggg aagttaggcc atattcgcac tgagcatgac gatgagcggc tcctgaaaga
     2281 accaaaattt gtaggttcat atatgattcc tgataacgaa gaccgagatg acaacaaaat
     2341 gtactttttc tttactgaga aggcgctgga ggcggagaac aacgcccaca cgatctacac
     2401 ccgagtgggg cggctgtgcg tgaatgacat gggaggacag agaatcctgg tgaacaagtg
     2461 gagcactttc cttaaagcgc ggctggtttg ctcagtgccg ggaatgaatg gaatcgacac
     2521 atactttgac gaactagagg atgtgttttt actgccgacc agagatccta agaatccagt
     2581 gatatttgga ctgtttaata ctaccagcaa tatatttaga ggccatgctg tatgtgtgta
     2641 tcacatgtca agtatccggg aagcctttaa tggcccatat gctcataaag aaggccctga
     2701 ataccactgg tcactatatg aaggaaaagt cccctaccca aggcctggtt cctgtgccag
     2761 caaagtaaac ggaggcaagt atggaaccac caaagattac cccgatgacg ccatccggtt
     2821 cgcaaggatg catcctctaa tgtatcagcc cataaaacct gttcataaaa aaccaatact
     2881 ggtaaaaaca gatggaaaat acaacctgag gcaacttgcc gtggatcggg tggaagcgga
     2941 ggatggccag tatgacgtct tatttattgg gacagacaca ggaattgtgc tgaaagtaat
     3001 cacaatttac aaccaagaaa cagagtggat ggaggaagtc attctagagg aacttcaaat
     3061 attcaaggat ccagccccta tcatttctat ggaaatttct tcaaagagac aacagcttta
     3121 cattggatca gcctctgctg tggcacaagt cagattccat cactgcgaca tgtatggcag
     3181 tgcttgtgct gactgctgcc tggctcgaga cccgtactgt gcctgggatg gcatatcctg
     3241 ctccaggtac tacccaacag gtgcacacgc aggcgcgcct ggatcttccg gaatcatccc
     3301 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     3361 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     3421 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     3481 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     3541 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     3601 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     3661 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     3721 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     3781 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     3841 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggggg
     3901 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     3961 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     4021 tgcccggtac gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtggc
     4081 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     4141 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     4201 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     4261 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     4321 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     4381 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     4441 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     4501 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     4561 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     4621 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     4681 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     4741 cgacgccgcg cacccgggtc atcatcacca tcaccattaa tctagagatc ttattaaagc
     4801 agaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4861 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4921 cttatcatgt ctggtcgact ctagctagac tcttccgctt cctcgctcac tgactcgctg
     4981 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5041 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5101 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5161 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5221 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5281 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5341 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5401 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5461 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5521 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5581 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5641 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5701 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5761 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5821 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6481 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6901 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6961 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggaggaatg tgtgtcagtt
     7021 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     7081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     7141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     7201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     7261 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7321 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc gtcccggaga
     7381 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     7441 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7501 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7561 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7621 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7681 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7741 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7801 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     7861 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     7921 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     7981 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc
     8041 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     8101 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     8161 aaaacgacgg ccagtgaatt
//
