LOCUS       sequence-131222-        9446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema5B protein, 
            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 9446)
  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 9446)
  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: Sema5b-AP-His
FEATURES             Location/Qualifiers
     source          1..9446
                     /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             6026..6043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6069..6190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6106..6125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6160..6179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6228..6245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6399..6987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6479..6498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7158..7949,7950..8018))
                     /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     7781..7800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8019..8123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8191..8209
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8247..8269)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8271..8291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8276..8606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8457..8592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8519..8538
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8743..8762
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8829..9284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8966..8987)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9178..9197
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9410..9432
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9424..9441
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9425..9441
                     /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 tggtagtccc
     1621 aggacccctg gctctctcgc tgttgctgtc cagcctcacc ctgctggtgt cccacctctc
     1681 cagctcccag gacattgcca gtgagtccag cagtgagcaa cagatgtgca cgcggaggga
     1741 gcaccccatc gtggcctttg aagatctgaa gccgtgggtc tttaacttca cccaccctgg
     1801 agtccgggat ttctcccagc ttgctcttga tccctcgagg aatcagctca tcgtgggagc
     1861 caggaactac ctcttcagac tcagcctcgc caatgtctcc ctccttcagg ctacagagtg
     1921 ggcctccagt gaggacacgc gccgctcctg ccagagcaaa gggaaaacgg aggaggagtg
     1981 tcagaactat gtacgagtcc tgattgtttc cggccggaag gtgttcatgt gcggtaccaa
     2041 tgccttttcc ccagtgtgct ccagcagaca ggtagggaac ctcagccgga ctattgagaa
     2101 gatcaacggt gtggcccggt gcccctatga cccacgccac aactctacag ccgtcatctc
     2161 ctctcagggg gagctctatg cagccacagt cattgacttc tccggtcggg acccagccat
     2221 ctaccgcagc ctgggcagtg ggccaccgct tcgcactgcc cagtataact ccaaatggct
     2281 caatgagcca aattttgtgg cagcctttga catcgggctg tttgcgtatt tcttccttcg
     2341 ggagaatgcc gtggagcatg attgtgggcg cactgtgtac tctcgggtgg cccgggtgtg
     2401 caagaatgat gtaggtggcc gtttcctgct ggaggacacg tggaccacat tcatgaaggc
     2461 ccggctcaac tgctcccgcc cgggagaggt ccccttctac tataatgagc tgcagagtgc
     2521 cttccatctg cccgagcagg acctcatcta tggcgtcttc accactaacg taaacagcat
     2581 tgcggcttct gctgtctgcg ccttcaacct cagtgccatc tccaaggctt tcaatggccc
     2641 atttcgttac caggaaaacc ccagggctgc ctggctccca atcgccaatc ccatccccaa
     2701 tttccagtgt ggcactctgc cggagactgg ccccaacgag aacctcacgg agcgcagcct
     2761 gcaggacgca cagcggctct tcctgatgag cgaagctgtg cagccagtga caccagagcc
     2821 ctgtgtcacc caggacagcg tccgcttctc acatctcgtg gtagaccttg tgcaagctaa
     2881 ggacacgctc tatcacgtac tctacatagg cacggagtcg ggtaccatcc tgaaagcgct
     2941 gtccacggcc agccgcagcc tccgtggctg ctacctggag gagctacatg tgctgcctcc
     3001 tgggcgcctt gaacctctgc ggagcctgcg catcctgcac agcgcgcgtg cgctcttcgt
     3061 gggattgagc gacagggtgc tgcgggtccc actggagagg tgttcggcct atcatagcca
     3121 gggggcatgc ctgggagcac gggacccata ctgcggctgg gatgggaagc ggcaactttg
     3181 cagcacgctt gaagacagtt ccaacatgag cctgtggatc cagaacatca caacctgtcc
     3241 tgtacgaaat gtgacacggg atgggggctt cggcccatgg tcaccatgga aaccgtgtga
     3301 gcacttagat ggagacaact cgggttcttg cctgtgccgg gccagatcct gtgactcccc
     3361 aaggcctcgc tgtgggggcc tcgagtgcct ggggccatcc atccatattg ccaattgctc
     3421 caggaatggg gcgtggaccg catggtcatc gtgggctcag tgcagcacgt cctgtgggat
     3481 cggcttccag gtccgtcagc gaagttgcag caacccggcg ccccgccatg ggggccgcat
     3541 ttgcgtgggc aagagccggg aggagcggtt ctgtaatgaa aatacacctt gcccagtgcc
     3601 catcttctgg gcttcctggg gttcctggag caagtgcagc aacaactgtg gaggcggcgt
     3661 gcagtcgcga cgccgttctt gcgagaatgg caactcgtgc ccgggttgcg gcgtggagtt
     3721 caagacctgc aaccccgagg cttgcccgga agtgcgacgc aacacaccct ggacgccctg
     3781 gctgcccgtg aacgtgaccc agggtggagc gcgccaggag cagcgattcc gcttcacctg
     3841 ccgcgcgccg ctgccggacc cgcacggtct gcagttcggc aagaggagga cagagaccag
     3901 gacttgcccg gcagacggca ccggagcctg cgacaccgac gccctggtgg aggatctcct
     3961 gcgcagcggg agcacgtcac cacacactct gaacggaggc tgggccacct ggggcccgtg
     4021 gtcatcctgc tcccgggact gcgagctggg cttccgcgtc cgcaagagaa cttgtaccaa
     4081 cccggagcct cgcaacgggg gcttgccctg cgtgggagac gctgcggagt accaagactg
     4141 caacccacag gcttgcccag tgcggggtgc ttggtcctgc tggaccgcat ggtcccagtg
     4201 ctcagcatcc tgcggtggtg gccactatca acgcacccgt tcctgcacca gccccgcccc
     4261 atccccaggt gaggacatct gcctcggcct gcacacggag gaggccctat gttcaacaca
     4321 ggcctgccca gaaggctggt cactgtggtc tgagtggggt gtctgcactg aggatggggc
     4381 ccagagccgg agccggagct gtgaggagct tctcccagga ccaggtgcct gtgttggcaa
     4441 cagcagccag agccggccct gcccctacag tgagattcct gtcatcctac ctgcttccag
     4501 tgtggaggag accaccagct gtggagggtt caatctcggc gcgcctggat cttccggaat
     4561 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     4621 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     4681 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     4741 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     4801 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     4861 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     4921 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     4981 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     5041 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     5101 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     5161 tgggggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     5221 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     5281 ccagggtgcc cggtacgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     5341 tgtggcccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     5401 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     5461 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     5521 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     5581 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     5641 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     5701 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     5761 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     5821 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     5881 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     5941 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     6001 caccaccgac gccgcgcacc cgggtcatca tcaccatcac cattaatcta gagatcttat
     6061 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6181 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac
     6241 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6301 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7201 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7261 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7321 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7381 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7441 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7501 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7561 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7621 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7681 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7741 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7801 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7861 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7921 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7981 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8101 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8161 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     8221 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg
     8281 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     8341 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     8401 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     8461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     8521 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     8581 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc
     8641 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     8701 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     8761 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     8821 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     8881 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     8941 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     9001 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     9061 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     9121 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     9181 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     9241 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     9301 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9361 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9421 cgttgtaaaa cgacggccag tgaatt
//
