LOCUS       sequence-137830-        9428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema5A 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 9428)
  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 9428)
  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: Sema5a-AP-His
FEATURES             Location/Qualifiers
     source          1..9428
                     /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             6008..6025
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6051..6172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6088..6107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6142..6161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6210..6227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6381..6969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6461..6480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7140..7931,7932..8000))
                     /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     7763..7782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8001..8105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8173..8191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8229..8251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8253..8273)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8258..8588
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8439..8574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8501..8520
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8725..8744
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8811..9266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8948..8969)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9160..9179
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9392..9414
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9406..9423
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9407..9423
                     /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 tgaagggagc
     1621 ctgcatcctt gcatggctgt tctcaagcct gggagtgtgg agacttgcta ggcccgagac
     1681 ccaggaccct gccaagtgcc agagagctga gcaccctgtc gtctcttaca aagaaattgg
     1741 cccctggtta cgggagttca gagccgagaa tgctgtggat ttctcgaggt taacatttga
     1801 cccaggacag aaagaacttg tcgtaggagc gagaaactat ctcttcagat tagagcttga
     1861 ggatctgtct ctcatccagg ctgtggagtg ggagtgtgac gaagccacca agaaggcctg
     1921 ttacagcaag ggcaaatcga aggaggaatg tcagaactac atacgagtgc tcttagttgg
     1981 tggggaccgg ctattcacct gtgggaccaa tgccttcaca cctgtctgta ccatccgctc
     2041 gttaagtaac ctgactgaga tccatgatca gatcagtggc atggcccgct gtccctacag
     2101 tccccagcac aattctaccg ccctactcac tgccagtggt gagctctatg ctgcaacagc
     2161 catggatttc ccagggcgag atccagccat ttaccgaagt ctgggcactt tgcctcctct
     2221 tcgaactgca cagtacaact ccaaatggct caatgaacca aactttgtgt cttcctacga
     2281 catcggaaat ttcacctact tcttcttccg agaaaacgcc gtggaacatg actgtgggaa
     2341 gacggtgttc tccagggctg cccgggtctg taaaaatgac attggagggc gatttcttct
     2401 ggaggacacc tggactacct tcatgaaggc tcgcctcaac tgctcccggc ctggtgaggt
     2461 gcccttctac tacaatgagc tgcagggtac tttcttcctg ccggagctgg atctgatcta
     2521 tggtatcttc accaccaatg taaacagcat cgctgcctct gctgtctgtg tcttcaacct
     2581 gagcgccatc tcacaggcct tcaatggacc cttcaagtac caagaaaact ctcgctcagc
     2641 ctggctgcct tatcccaacc ccaaccccaa ttttcagtgc ggcaccatgg accagggcct
     2701 gtacgtaaac ctgacggaaa gaaacctaca ggatgctcag aagttcattc tgatgcatga
     2761 agtggtgcag ccagtgacca cagtgccctc tttcatggag gacaacagcc gcttctccca
     2821 cttagctgtt gatgttgtgc aaggcaggga gactctggtc cacatcattt atctggccac
     2881 agattatggc accattaaga aagtacgagc tcccctgagt cagagctcag gcagctgttt
     2941 gctggaagag attgagcttt tcccagagag gaggagtgag cccatcagga gcctgcagat
     3001 cctccacagc cagagtgtcc tatttgtggg actacaggag catgtggcca agatccccct
     3061 gaagaggtgc cacttccatc aaacacgcag tgcctgcatt ggcgctcagg acccttactg
     3121 tggctgggat gcggtgatga agaaatgcac cagcctggag gagagtctga gcatgaccca
     3181 gtgggatcag agcatcccca cctgtccgac cagaaatctc actgtagatg ggagctttgg
     3241 cccatggtca ccgtggacac cctgtacaca cactgatggc actgctgtgg gctcctgcct
     3301 ctgccggtcc cgctcctgtg acagcccagc tcctcagtgt ggtggttggc agtgtgaggg
     3361 ccctagaatg gagatcacca actgttccag gaatggaggc tggactccct ggacctcctg
     3421 gtctccgtgc agcacgacct gtggcattgg cttccaagtg cggcagcgat cctgcagcaa
     3481 ccccacgccc aggcatggcg ggcgcgtgtg cgtgggccag aacagggagg aaagatactg
     3541 caatgaacat ttgctctgcc caccacacgt gttctggaca ggctggggac cttgggaacg
     3601 gtgcacagcc cagtgcgggg gtggcattca agcccgccgg aggacctgtg aaaatgggcc
     3661 tgactgtgca ggatgcaatg tggaatacca gccttgtaac accaatgcat gcccggagct
     3721 gaagaagacc acaccctgga caccctggac ccctgtcaac atctctgaca atggaggtca
     3781 ctatgagcag cgtttccgct atacctgtaa agctcgcctg ccagatccaa atttgctgga
     3841 agtaggaaga cagaggatag aaatgcggta ctgttccagc gatggaacca gtggctgctc
     3901 cacagacgga ctttctggag actttctaag agctgggaga tactctgctc atacagtcaa
     3961 tggggcatgg tcagcctgga cttcctggtc acagtgcagc agagactgca gcaggggcat
     4021 tcggaaccgg aagcgtgttt gcaacaaccc agaacccaag tttgggggca tgccatgcct
     4081 tggcccatcg ctggagttcc aggaatgcaa cattttacct tgtccagtgg atggtgtgtg
     4141 gtcttgctgg tcatcctggt ctaaatgttc agcaacctgt ggaggtggcc actacatgag
     4201 aacccgttct tgttcgaatc cagccccagc atatggaggg gacatctgcc tgggactgca
     4261 cacagaagag gcactctgca acacacagac ctgcccagaa agctggtcag agtggtcaga
     4321 ctggtctgtg tgcgatgcat ctggtaccca ggtccgtgct cggcagtgca tccttctgtt
     4381 tccagtgggc agccaatgtt ctggaaatac tacagagagc cggccttgtg tatttgactc
     4441 taatttcatc ccagaagtat ctgtggcaag atccagcagc gtagaagaaa aaaggtgtgg
     4501 agagttcaac atgttccacg gcgcgcctgg atcttccgga atcatcccag ttgaggagga
     4561 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     4621 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     4681 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     4741 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     4801 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     4861 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     4921 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     4981 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     5041 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     5101 tacgcagctc atctccaaca tggacattga cgtgatccta ggtgggggcc gaaagtacat
     5161 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     5221 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     5281 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     5341 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     5401 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     5461 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     5521 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     5581 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     5641 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     5701 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     5761 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     5821 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     5881 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     5941 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     6001 cccgggtcat catcaccatc accattaatc tagagatctt attaaagcag aacttgttta
     6061 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6121 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6181 ggtcgactct agctagactc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6781 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8101 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8161 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     8221 ggtgaaaacc tctgacacat gcagctcccg gaggaatgtg tgtcagttag ggtgtggaaa
     8281 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     8341 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     8401 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     8461 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     8521 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     8581 tttgcaaaaa gctcccggga gcttgtatat ccattttcgt cccggagacg gtcacagctt
     8641 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     8701 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     8761 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     8821 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     8881 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     8941 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     9001 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     9061 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     9121 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     9181 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     9241 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     9301 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     9361 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     9421 agtgaatt
//
