LOCUS       sequence-131226-        7973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema6B protein (truncated 
            at extracellular domain cleavage site; 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 7973)
  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 7973)
  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: Sema6b(S)-AP-His
FEATURES             Location/Qualifiers
     source          1..7973
                     /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             4553..4570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4596..4717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4633..4652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4687..4706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4755..4772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4926..5514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5006..5025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5685..6476,6477..6545))
                     /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     6308..6327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6546..6650)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6718..6736
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6774..6796)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6798..6818)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6803..7133
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6984..7119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7046..7065
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7270..7289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7356..7811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7493..7514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7705..7724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7937..7959
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7951..7968
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7952..7968
                     /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 tgtggacccc
     1621 gcgagtgccc cctccacgcc cggccctgtc gtttttcctg ctgttgctcc tgggggtcac
     1681 ctatggcctt ttcccagaag aaccacctcc actcagcgtg gctcccaggg actacttgag
     1741 ccactatccc gtgttcgtgg gcagcgggcc tggtcgtctg accgctgcgg agggtgctga
     1801 ggacctcaac atccagagag tgctacgtgt taacaggaca ctgttcattg gggacagaga
     1861 taacctgtac caagtagaac tggagccatc cacttccacg gagctgcgat atcagaggaa
     1921 gctgacctgg cgctccaacc ccagtgacat cgatgtgtgt cgaatgaagg gcaagcaaga
     1981 gggtgagtgt cggaactttg tcaaggtgct cctgcttcgc gatgagtcca cgctgttcgt
     2041 gtgcggctcc aatgcgttca atcccatctg cgccaattac agtatggaca cactgcagct
     2101 tcttggtgac agcatcagcg gtatggcccg ctgcccctat gaccccaaac atgccaatgt
     2161 cgccctcttc tcagatggga tgctcttcac agccacggtg actgacttcc tagccatcga
     2221 cgctgttatc taccgcagcc ttggggaccg gcccacactg cgcacagtaa agcacgactc
     2281 caagtggttt aaagagccgt actttgtgca tgcagtggag tggggaagcc acgtctattt
     2341 cttcttccgg gagatcgcca tggagtttaa ctacctggaa aaggtggtgg tgtctcgagt
     2401 ggcccgtgtg tgcaagaatg acgtgggcgg ctcgccacga gtgctggaga agcagtggac
     2461 ttccttcctg aaggcccggc tcaactgctc agtgccaggg gattcacact tctacttcaa
     2521 cgtcctgcag gctgtgacgg gcgtggtgag cctgggtggt cgtccggtga tcctcgctgt
     2581 cttctcaact cctagcaaca gcatccccgg ctcagccgtc tgcgcctttg acatgaacca
     2641 agtggctgct gtgtttgaag gccgcttccg ggaacagaag tcgcctgagt cgatctggac
     2701 accagtgcct gaagaccaag tgccacggcc caggcctggg tgctgcgcgg cacctggcat
     2761 gcagtacaac gcgtccagtg ccctccctga cgagatcctc aactttgtaa agacccaccc
     2821 actgatggat gaggcagtac cttccctggg tcactcgccg tggattgtga gaactctgat
     2881 gcggcaccag ctgacccgag tggctgtgga tgtgggtgcg ggtccatggg gcaatcagac
     2941 aatagtcttc cttggctccg aggctggcac agtcctcaaa tttcttgtga agcccaacgc
     3001 cagtgtctca gggaccacag ggcccagcat ctttttggag gagtttgaga cctaccggcc
     3061 agacggcgcg cctggatctt ccggaatcat cccagttgag gaggagaacc cggacttctg
     3121 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     3181 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     3241 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     3301 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     3361 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     3421 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     3481 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     3541 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     3601 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     3661 caacatggac attgacgtga tcctaggtgg gggccgaaag tacatgtttc gcatgggaac
     3721 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     3781 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tacgtgtgga accgcactga
     3841 gctcatgcag gcttccctgg acccgtctgt ggcccatctc atgggtctct ttgagcctgg
     3901 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     3961 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     4021 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     4081 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     4141 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     4201 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     4261 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     4321 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     4381 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     4441 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     4501 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gtcatcatca
     4561 ccatcaccat taatctagag atcttattaa agcagaactt gtttattgca gcttataatg
     4621 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4681 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagcta
     4741 gactcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5101 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5341 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5761 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5821 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5881 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5941 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6001 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6061 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6661 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6721 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6781 cacatgcagc tcccggagga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6841 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6961 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7021 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     7081 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     7141 cgggagcttg tatatccatt ttcgtcccgg agacggtcac agcttgtctg taagcggatg
     7201 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     7261 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     7321 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     7381 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7441 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     7501 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     7561 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     7621 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     7681 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     7741 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     7801 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     7861 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     7921 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
