LOCUS       sequence-137721-        8459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema6A, isoform a 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 8459)
  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 8459)
  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: Sema6a.a-AP-His
FEATURES             Location/Qualifiers
     source          1..8459
                     /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             5039..5056
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5082..5203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5119..5138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5173..5192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5241..5258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5412..6000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5492..5511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6171..6962,6963..7031))
                     /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     6794..6813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7032..7136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7204..7222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7260..7282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7284..7304)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7289..7619
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7470..7605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7532..7551
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7756..7775
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7842..8297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7979..8000)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8191..8210
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8423..8445
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8437..8454
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8438..8454
                     /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 tgcggccagc
     1621 agccttactg ctgtgtctca cactgctaca ctgcgctggg gctggtttcc cagaagattc
     1681 cgagccaatc agtatttcgc atggcaacta tacaaaacag tatccggtgt ttgtgggcca
     1741 caagccagga cggaacacca cgcagaggca caggctggac atccagatga tcatgatcat
     1801 gaacagaacc ctctacgttg ctgctcgaga ccatatttat actgttgata tagacacatc
     1861 ccacacagaa gaaatttact gtagcaaaaa actgacatgg aaatctagac aggctgacgt
     1921 agacacatgc aggatgaagg ggaaacataa ggatgaatgt cacaacttca ttaaagttct
     1981 tctcaagaag aatgatgata cgctgtttgt ctgtggaacc aatgccttca acccttcctg
     2041 cagaaactac agggtcgata ccttggaaac ttttggggat gaatttagcg gaatggccag
     2101 atgcccttat gatgccaaac atgccaacat cgctctgttt gcagatggaa aactctactc
     2161 ggctacagtg actgactttc tggccattga tgcggtcatt tacaggagcc tcggagacag
     2221 ccctaccctc aggactgtca agcatgattc aaagtggttg aaagagccgt actttgtcca
     2281 agccgtggat tatggggact atatctactt cttcttcaga gaaattgcag tagaatacaa
     2341 cactatgggg aaggttgttt tccctagggt ggctcaggtc tgtaagaatg acatgggagg
     2401 gtctcagaga gtcctggaga agcagtggac atctttcctg aaggctcgcc tgaactgctc
     2461 ggtgcctgga gactctcatt tttatttcaa tatactccag gcagttacag atgtgattcg
     2521 cattaatggc cgtgatgttg tcttggcaac cttttccaca ccttataaca gcatcccagg
     2581 ttctgcagtc tgtgcctatg acatgcttga cattgctaat gttttcactg ggaggttcaa
     2641 ggaacagaaa tcacctgact ctacctggac acccgttcca gacgaacgag tccctaagcc
     2701 caggccaggc tgttgtgctg gatcatcctc tttagaaaaa tatgcaacct ccaatgagtt
     2761 tcccgatgat accctgaact tcattaagac gcatccactc atggacgagg cagtgccttc
     2821 catcatcaac agaccttggt tcctgagaac aatggtcaga taccgcctga ccaaaattgc
     2881 agtagacaac gctgccgggc catatcagaa tcacactgtg gttttcctgg gatcagaaaa
     2941 gggaatcatc ctgaagttct tggccaggat aggaagcagt ggtttcctaa atggcagcct
     3001 tttcctggag gagatgaatg tttacaaccc agaaaagtgc agctatgatg gtgtagaaga
     3061 caaaaggatc atgggcatgc agctcgacag agcgagtggc tcactctatg ttgcattctc
     3121 tacttgtgtg atcaaggtgc ctcttggccg ctgtgagcga catgggaagt gtaaaaaaac
     3181 ctgcatcgcc tccagagacc cgtattgtgg gtgggtaagg gaaagtggtt cctgtgccca
     3241 tctgtcaccc cttagcagac tgacatttga gcaggacatt gagcgtggca atacggacgg
     3301 cctaggagac tgtcacaatt ccttcgtggc actgaatggg cacgccagtt ccctctatcc
     3361 cagcaccact acgtcagatt cggcatcccg agacgggtat gagtctaggg gaggcatgct
     3421 ggactggaac gacctgctcg aggcacctgg cagcacagac cctttggggg cagtgtcctc
     3481 tcataaccac caggacaaga agggagtgat tcgggaaagt tacctcaaaa gcaacgacca
     3541 gcttgttcct ggcgcgcctg gatcttccgg aatcatccca gttgaggagg agaacccgga
     3601 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     3661 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     3721 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     3781 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     3841 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     3901 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     3961 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     4021 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     4081 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     4141 catctccaac atggacattg acgtgatcct aggtgggggc cgaaagtaca tgtttcgcat
     4201 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     4261 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     4321 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     4381 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     4441 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     4501 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     4561 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     4621 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     4681 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     4741 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     4801 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     4861 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     4921 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     4981 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtca
     5041 tcatcaccat caccattaat ctagagatct tattaaagca gaacttgttt attgcagctt
     5101 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggtcgactc
     5221 tagctagact cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5281 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5341 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7201 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     7261 ctctgacaca tgcagctccc ggaggaatgt gtgtcagtta gggtgtggaa agtccccagg
     7321 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     7381 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7441 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     7501 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     7561 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     7621 agctcccggg agcttgtata tccattttcg tcccggagac ggtcacagct tgtctgtaag
     7681 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7741 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7801 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     7861 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     7921 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     7981 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     8041 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     8101 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     8161 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     8221 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     8281 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     8341 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     8401 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatt
//
