LOCUS       sequence-131205-        8786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3E protein (truncated at cleavage site P3; ie, 
            long), 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 8786)
  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 8786)
  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(L)-AP-His
FEATURES             Location/Qualifiers
     source          1..8786
                     /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             5366..5383
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5409..5530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5500..5519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5568..5585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5739..6327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5819..5838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6498..7289,7290..7358))
                     /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     7121..7140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7359..7463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7531..7549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7587..7609)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7611..7631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7616..7946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7797..7932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7859..7878
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8083..8102
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8169..8624)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8306..8327)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8518..8537
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8750..8772
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8764..8781
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8765..8781
                     /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 aaagaggagg ttccgcaggc aggacgttcg
     3301 gcatggcaac gccgcccaac agtgctttgg acagcaattt gttggagacg cgttggacag
     3361 gactgaagag aggctggctt atggcataga gagcaacagt actctgttgg aatgcacccc
     3421 gcgatcacta caagcaaaag tcatctggtt tgtacagaag ggacgcgacg taagaaaaga
     3481 agaggtgaag acggatgaca gagttgtcaa gatggacttg ggcttgctct tcctcagagt
     3541 acgcaagtca gatgcaggga cctatttttg ccagacagta gaacacaatt ttgtccatac
     3601 tgtgcgtaaa atcaccttgg aggtggtcga agagcataaa gtggagggca tgtttcataa
     3661 ggaccatgaa gaggaaagac atcacaagat gccctgccct cccttaagcg gtatgtctca
     3721 ggggacaaaa ccgtggtaca aggaattctt gcagctgatt ggctacagca acttccagag
     3781 agtggaagaa tactgcgaaa aggtgtggtg tacagataag aagaggaaaa agcttaaaat
     3841 gtctccctcc aagtggaagt atgccaaccc ccaggaaggc gcgcctggat cttccggaat
     3901 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     3961 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     4021 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     4081 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     4141 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     4201 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     4261 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     4321 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     4381 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     4441 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     4501 tgggggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     4561 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     4621 ccagggtgcc cggtacgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     4681 tgtggcccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     4741 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     4801 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     4861 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     4921 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     4981 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     5041 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     5101 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     5161 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     5221 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     5281 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     5341 caccaccgac gccgcgcacc cgggtcatca tcaccatcac cattaatcta gagatcttat
     5401 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5521 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac
     5581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7501 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7561 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg
     7621 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     7681 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     7741 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     7801 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     7861 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     7921 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc
     7981 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     8041 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     8101 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     8161 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     8221 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     8281 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     8341 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     8401 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     8461 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     8521 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     8581 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     8641 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8701 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8761 cgttgtaaaa cgacggccag tgaatt
//
