LOCUS       sequence-135522-        8162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3B 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 8162)
  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 8162)
  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: Sema3b(S)-AP-His
FEATURES             Location/Qualifiers
     source          1..8162
                     /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             4742..4759
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4785..4906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4822..4841)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4876..4895
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4944..4961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5115..5703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5195..5214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5874..6665,6666..6734))
                     /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     6497..6516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6735..6839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6907..6925
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6963..6985)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6987..7007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6992..7322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7173..7308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7235..7254
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7459..7478
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7545..8000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7682..7703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7894..7913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8126..8148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8140..8157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8141..8157
                     /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 tggggcgggc
     1621 tgaggccgcc gccatgatcc caggcctggc ccttctctgg gtagcagggc taggggatac
     1681 tgcccctaac cttccccgcc ttcggctctc ctttcaagaa ttacaggccc ggcatggtgt
     1741 ccgaaccttc aggctggagc ggacctgctg ttatgaagcc ttgctggtgg atgaggagcg
     1801 tggacgcctg tttgtgggtg ctgagaacca cgtggcttcc ctcagcctgg acaacatcag
     1861 caagcgagcc aagaagttgg cctggcccgc ccccgtggaa tggcgtgaag aatgcaactg
     1921 ggcagggaag gacattggta ccgagtgcat gaacttcatg aagctgctgc acgcctacaa
     1981 ccacacccac ttgctggcct gtggcacagg ggctttccac ccaacctgcg cctttgtgga
     2041 ggtgggccac cggctggagg aacccatgct tcaactggac cggaggaaac ttgaggacgg
     2101 caaggggaag actccttatg acccaaggca tcgggctgcc tcggtgctgg tgggggaaga
     2161 gctgtattct ggggtggcag cagaccttat gggccgggac tttaccatct ttcgaagcct
     2221 tggtcagaat ccgagtctcc gaacagagcc ccatgattcc cgctggctca atgaacccaa
     2281 gtttgtcaag gtcttttgga tcccagagag tgagaaccct gatgacgata aaatctattt
     2341 cttcttccgc gagtccgctg tggaagcagc accagcaatg gggcgcatgt ctgtgtctcg
     2401 tgttggccag atctgcagga atgacctggg tggccagcgg agcttggtca acaaatggac
     2461 cacatttctg aaggcgcggc ttgtgtgctc agtacctgga gttgagggtg acacccactt
     2521 tgaccaactt caggatgttt tccttctgtc ctcccgagac cgccagacac ctcttctcta
     2581 tgctgtcttc tccacctcca gtggtgtctt ccagggctct gctgtgtgcg tgtacagcat
     2641 gaacgatgtg cgccgagcct tcttgggacc ttttgctcac aaagaggggc ctacacacca
     2701 gtgggtgtcc taccagggtc gtgtccccta cccaagacct ggcatgtgcc ccagcaagac
     2761 ctttggcacc ttcagctcca ccaaggactt cccagatgac gttatccagt ttgctcggaa
     2821 ccaccctctc atgtacaacc cagtcctgcc catggggggg cgccctctct tcctacaagt
     2881 gggagctggg tacaccttca cccaaatcgc cgcagaccga gtagcagctg ccgatggaca
     2941 ctacgatgtt ctcttcattg gtacagatgt gggcacagtg ctgaaagtga tctcagtccc
     3001 caaaggcagc cgacctaatt ctgaaggact tctcctggaa gagctgcagg tgttcgagga
     3061 ctctgccgct atcaccagca tgcaaatctc ctctaaaagg caacaactct acgtagcatc
     3121 gcgcagcgca gtggcccaga ttgctttgca tcgctgcact gccctaggcc gcgcctgcgc
     3181 agaatgctgc ttggcccgtg atccttactg cgcctgggat ggatcagctt gcacacgctt
     3241 ccagcctacg gccggcgcgc ctggatcttc cggaatcatc ccagttgagg aggagaaccc
     3301 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     3361 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     3421 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     3481 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     3541 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     3601 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     3661 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     3721 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     3781 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     3841 gctcatctcc aacatggaca ttgacgtgat cctaggtggg ggccgaaagt acatgtttcg
     3901 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     3961 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
     4021 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     4081 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     4141 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     4201 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     4261 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     4321 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     4381 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     4441 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     4501 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     4561 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     4621 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     4681 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     4741 tcatcatcac catcaccatt aatctagaga tcttattaaa gcagaacttg tttattgcag
     4801 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4861 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggtcga
     4921 ctctagctag actcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5521 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6181 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6481 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6841 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6901 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6961 aacctctgac acatgcagct cccggaggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     7021 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     7081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     7141 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7201 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7261 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     7321 aaaagctccc gggagcttgt atatccattt tcgtcccgga gacggtcaca gcttgtctgt
     7381 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     7441 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     7501 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     7561 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     7621 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     7681 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     7741 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     7801 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     7861 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     7921 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     7981 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     8041 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     8101 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     8161 tt
//
