LOCUS       sequence-136383-        8699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sema3B 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 8699)
  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 8699)
  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(L)-AP-His
FEATURES             Location/Qualifiers
     source          1..8699
                     /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             5279..5296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5322..5443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5359..5378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5413..5432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5481..5498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5652..6240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5732..5751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6411..7202,7203..7271))
                     /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     7034..7053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7272..7376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7444..7462
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7500..7522)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7524..7544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7529..7859
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7710..7845
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7772..7791
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7996..8015
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8082..8537)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8219..8240)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8431..8450
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8663..8685
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8677..8694
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8678..8694
                     /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 aagaagctgg cctggcccgc ccccgtggaa tggcgtgaag aatgcaactg
     1921 ggcagggaag gacattggta ccgagtgcat gaacttcgtg aagctgctgc acacctacaa
     1981 ccacacccac ttgctggcct gtggcacagg ggctttccac ccaacctgtg cctttgtgga
     2041 ggtgggccac cggctggagg aacccatgct tcaactggac cggaggaaac ttgaggacgg
     2101 caaggggaag actccttatg acccaaggca tcgggctgcc tcggtgctgg tgggggaaga
     2161 actgtattct ggggtgacag 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 acatagcatc
     3121 gcgcagcgca gtggcccaga ttgctttgca tcgctgcact gccctaggcc gcgcctgcgc
     3181 agaatgctgc ttggcccgtg atccttactg cgcctgggat ggatcagctt gcacacgctt
     3241 ccagcctacg gccaagagac ggttccggag gcaagacata aggaatggcg accccagcac
     3301 cctatgctct ggagactctt ctcactctgt gctgctggag aagaaggtgt tgggtgtgga
     3361 gagcggcagc gcgtttctgg agtgtgagcc ccgctcgctc caggcgcatg tgcagtggac
     3421 cttccaaggt gcaggggagg cagctcacac ccaggtgctg gctgaggaga gagtagagcg
     3481 cactgcgcgg gggctgctgt tgcgggggct gcggcgccag gactctggcg tgtatctttg
     3541 cgtcgcggtt gaacaaggct tttcacaacc actgcgtcgc ctggtgctgc atgtgttgag
     3601 tgcggcgcag gctgaacgac tggcacgggc agaggaagca gccgctcctg cacctcctgg
     3661 ccctaaactc tggtaccggg actttctgca gttggtggag ccaggcggtg gcggaggtgc
     3721 aaactccctg cgaatgtgcc gcccgcagcc cgggcaccac tctgtggcag cagattcacg
     3781 tcgtaagggt ggcgcgcctg gatcttccgg aatcatccca gttgaggagg agaacccgga
     3841 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     3901 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     3961 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     4021 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     4081 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     4141 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     4201 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     4261 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     4321 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     4381 catctccaac atggacattg acgtgatcct aggtgggggc cgaaagtaca tgtttcgcat
     4441 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     4501 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     4561 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     4621 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     4681 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     4741 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     4801 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     4861 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     4921 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     4981 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     5041 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     5101 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     5161 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     5221 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtca
     5281 tcatcaccat caccattaat ctagagatct tattaaagca gaacttgttt attgcagctt
     5341 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5401 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggtcgactc
     5461 tagctagact cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5521 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5581 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6061 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6301 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7381 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7441 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     7501 ctctgacaca tgcagctccc ggaggaatgt gtgtcagtta gggtgtggaa agtccccagg
     7561 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     7621 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7681 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     7741 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     7801 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     7861 agctcccggg agcttgtata tccattttcg tcccggagac ggtcacagct tgtctgtaag
     7921 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7981 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     8041 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     8101 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     8161 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     8221 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     8281 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     8341 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     8401 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     8461 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     8521 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     8581 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     8641 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatt
//
