LOCUS       sequence-131231-        8363 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the Sema6D, isoform 5 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 8363)
  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 8363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: Sema6d.5-AP-His
FEATURES             Location/Qualifiers
     source          1..8363
                     /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             4943..4960
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4986..5107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5023..5042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5077..5096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5145..5162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5316..5904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5396..5415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6075..6866,6867..6935))
                     /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     6698..6717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6936..7040)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7108..7126
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7164..7186)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7188..7208)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7193..7523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7374..7509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7436..7455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7660..7679
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7746..8201)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7883..7904)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8095..8114
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8327..8349
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8341..8358
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8342..8358
                     /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 tggggttcct
     1621 tctgctttgg ttctgcgtgc tgttccttct ggtctccagg ttacgggcgg tcagcttccc
     1681 agaagacgat gagcccctca acacggttga ctatcactat tcaaggcaat atccggtttt
     1741 tagaggacgc ccttcaggca acgaatcgca gcacaggctg gactttcagc tgatgttgaa
     1801 aattcgagac acactttata ttgctggcag ggatcaagtc tatacagtga acttaaatga
     1861 aatcccccaa acagaggtga taccaagcaa gaagctgacg tggaggtcca gacagcagga
     1921 tcgagaaaat tgtgctatga aaggcaagca taaagatgaa tgccacaact tcatcaaagt
     1981 ctttgtccca agaaatgatg agatggtttt tgtctgtggt accaatgctt tcaacccgat
     2041 gtgcagatac tataggttga gaacgttaga gtatgatggg gaagaaatta gtggcctggc
     2101 acgatgcccg tttgatgccc gacaaaccaa tgtcgccctc tttgctgatg gaaaactcta
     2161 ttctgccaca gtggctgatt tcctggccag tgatgctgtc atttacagaa gcatgggaga
     2221 tggatctgcc cttcgcacaa taaaatacga ttccaagtgg atcaaagaac cacacttcct
     2281 tcatgccata gaatatggaa actatgtcta tttcttcttc agagaaatcg ccgtggaaca
     2341 taataactta ggcaaggctg tgtattcccg cgtggctcgc atttgtaaaa acgacatggg
     2401 tggctcacag cgggtcctgg agaaacactg gacttccttc cttaaggctc ggctgaactg
     2461 ctccgttcct ggagattcct ttttctactt cgacgtcctg cagtctataa cagacataat
     2521 ccaaatcaat ggcatcccca ctgtggttgg ggtcttcacc acacagctca acagcattcc
     2581 tggttctgca gtctgtgcct ttagcatgga cgacattgag aaagtgttca aagggcggtt
     2641 caaagagcag aaaaccccag actctgtttg gacagcagtt cccgaagaca aagtaccaaa
     2701 accaaggcct ggctgttgtg ccaaacacgg cctcgcagaa gcttacaaga cctccatcga
     2761 ctttccagat gacaccctgg ctttcatcaa gtcccacccg ctgatggact ctgccgtccc
     2821 acccattgcc gatgagccct ggttcacaaa gacacgggtc aggtacaggt tgacagccat
     2881 cgaagtggac cgttcagcag ggccatacca aaactacaca gtcatctttg ttggctctga
     2941 agctggcgtg gtacttaaag ttttggcaaa gaccagtcct ttctctctga atgacagtgt
     3001 attactcgaa gagattgaag cttataaccc agccaagtgc agcgccgaga gtgaggagga
     3061 cagaaaggtg gtctcattac agctggacaa ggatcaccat gctttatacg tggccttctc
     3121 tagctgcgtg gtccgcatcc ccctcagccg ctgtgagcgc tacggatcgt gtaaaaagtc
     3181 ttgcattgca tcacgtgacc cgtactgtgg ttggttaagc cagggagttt gtgagagagt
     3241 gaccctaggg atgctgctgt taaccgaaga cttctttgct ttccataacc acagccctgg
     3301 aggatatgag caggacacgg agtacggcaa cacagcccac ctaggggact gccacgaaag
     3361 tttgcctcct tcaactacac cagattacaa aatatttggc ggtccaacat ctggtgtacg
     3421 gtgggaagtc cagtctggag aatccaatca gatggtcact agtggatctt ccggaatcat
     3481 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     3541 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     3601 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     3661 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     3721 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     3781 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     3841 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     3901 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     3961 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     4021 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     4081 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     4141 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     4201 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     4261 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     4321 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     4381 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     4441 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     4501 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     4561 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     4621 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     4681 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     4741 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     4801 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     4861 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     4921 caccgacgcc gcgcacccgg gtcatcatca ccatcaccat taatctagag atcttattaa
     4981 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5041 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5101 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg
     5161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6121 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6181 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6241 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6301 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6361 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6421 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6481 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6541 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6601 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6661 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6721 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6781 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6841 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6901 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7081 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     7141 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca
     7201 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     7261 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     7321 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     7381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     7441 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     7501 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg
     7561 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7621 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7681 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7741 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     7801 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     7861 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     7921 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7981 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     8041 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     8101 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     8161 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     8221 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     8281 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     8341 tgtaaaacga cggccagtga att
//
