LOCUS       sequence-135624-        8216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3D 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 8216)
  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 8216)
  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(S)-AP-His
FEATURES             Location/Qualifiers
     source          1..8216
                     /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             4796..4813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4839..4960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4876..4895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4930..4949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4998..5015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5169..5757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5249..5268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5928..6719,6720..6788))
                     /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     6551..6570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6789..6893)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6961..6979
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7017..7039)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7041..7061)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7046..7376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7227..7362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7289..7308
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7513..7532
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7599..8054)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7736..7757)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7948..7967
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8180..8202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8194..8211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8195..8211
                     /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 caatacagaa tgtgcaaatt tcatccgtgt gcttcaaccc tataataaga ctcacgttta
     2041 cgtgtgtgga actggagcgt ttcatccgct gtgtgggtac atcgatctcg 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 cacgtctggc gcgcctggat cttccggaat catcccagtt gaggaggaga acccggactt
     3361 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     3421 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     3481 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     3541 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     3601 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     3661 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     3721 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     3781 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     3841 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     3901 ctccaacatg gacattgacg tgatcctagg tgggggccga aagtacatgt ttcgcatggg
     3961 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     4021 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     4081 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     4141 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     4201 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     4261 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     4321 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     4381 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     4441 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     4501 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     4561 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     4621 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     4681 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     4741 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggtcatca
     4801 tcaccatcac cattaatcta gagatcttat taaagcagaa cttgtttatt gcagcttata
     4861 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4921 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg tcgactctag
     4981 ctagactctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5041 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5101 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5341 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5581 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5641 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5881 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5941 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6001 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6061 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6121 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6181 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6241 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6301 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6361 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6421 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6481 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6541 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6601 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6661 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6721 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6781 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6901 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6961 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     7021 tgacacatgc agctcccgga ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     7081 cccaggcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     7141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     7201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     7261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     7321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     7381 tcccgggagc ttgtatatcc attttcgtcc cggagacggt cacagcttgt ctgtaagcgg
     7441 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     7501 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     7561 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     7621 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     7681 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     7741 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     7801 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     7861 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     7921 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     7981 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     8041 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     8101 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     8161 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaatt
//
