LOCUS       sequence-131224-        8294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema6A, isoform z 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 8294)
  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 8294)
  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.z-AP-His
FEATURES             Location/Qualifiers
     source          1..8294
                     /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             4874..4891
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4917..5038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4954..4973)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5008..5027
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5076..5093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5247..5835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5327..5346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6006..6797,6798..6866))
                     /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     6629..6648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6867..6971)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7039..7057
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7095..7117)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7119..7139)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7124..7454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7305..7440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7367..7386
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7591..7610
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7677..8132)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7814..7835)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8026..8045
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8258..8280
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8272..8289
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8273..8289
                     /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 actgaatgga gtgattcggg aaagttacct
     3361 caaaagcaac gaccagcttg ttcctggcgc gcctggatct tccggaatca tcccagttga
     3421 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     3481 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     3541 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     3601 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     3661 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     3721 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     3781 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     3841 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     3901 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     3961 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg ggggccgaaa
     4021 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     4081 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     4141 gtacgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tggcccatct
     4201 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     4261 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     4321 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     4381 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     4441 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     4501 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     4561 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     4621 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     4681 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     4741 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     4801 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     4861 cgcgcacccg ggtcatcatc accatcacca ttaatctaga gatcttatta aagcagaact
     4921 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4981 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5041 atgtctggtc gactctagct agactcttcc gcttcctcgc tcactgactc gctgcgctcg
     5101 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5161 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5761 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5821 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5881 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6001 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6061 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6121 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6961 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     7021 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     7081 gatgacggtg aaaacctctg acacatgcag ctcccggagg aatgtgtgtc agttagggtg
     7141 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     7201 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     7261 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     7321 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7381 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7441 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcgtcccg gagacggtca
     7501 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     7561 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     7621 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     7681 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7741 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7801 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7861 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7921 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7981 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     8041 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8101 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     8161 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     8221 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8281 acggccagtg aatt
//
