LOCUS       sequence-131144-        8093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the LRRC4 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 8093)
  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 8093)
  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: Lrrc4-AP-His
FEATURES             Location/Qualifiers
     source          1..8093
                     /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             4673..4690
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4716..4837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4753..4772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4807..4826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4875..4892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5046..5634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5126..5145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5805..6596,6597..6665))
                     /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     6428..6447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6666..6770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6838..6856
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6894..6916)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6918..6938)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6923..7253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7104..7239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7166..7185
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7390..7409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7476..7931)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7613..7634)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7825..7844
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8057..8079
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8071..8088
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8072..8088
                     /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 tgaagctctt
     1621 gtggcaggta actgtgcacc acacctggaa tgccgtcctg cttcccgtcg tctacctcac
     1681 ggcgcaagtg tggattctgt gtgcagccat cgctgctgcc gcttcagccg ggccccaaaa
     1741 ctgcccctcc gtctgttcct gcagtaacca gttcagcaag gtggtgtgca cccgccgggg
     1801 actctctgag gtcccacagg gtattccttc caacacccgg tatctcaacc tcatggaaaa
     1861 caacatccag atgattcagg ccgacacctt caggcacctc catcacctgg aggtcctgca
     1921 gctgggcagg aactccatca ggcagataga ggtgggggcc ttcaatggcc tcgccagtct
     1981 caacactctg gaactgttcg acaactggct gacagtcatc cccagtgggg cctttgagta
     2041 cctgtccaaa ctgcgggagc tctggctgcg aaacaatccc attgagagca tcccttctta
     2101 tgccttcaac cgggtgccct ccctcatgcg tctggacttg ggggaactca agaagctgga
     2161 gtatatctct gagggggctt ttgaggggct gttcaacctc aagtacctga acttgggcat
     2221 gtgcaacatt aaggatatgc ccaatcttac acccctggtg gggctcgagg agcttgagat
     2281 gtcagggaac cactttcctg agatcaggcc tgggtccttc catggcctaa gctccctcaa
     2341 aaaactgtgg gtgatgaact cacaggtcag cctgattgag cggaatgctt ttgacggact
     2401 ggcttccctc gtggaactca acttagccca caataacctc tcatctttgc cccatgacct
     2461 cttcacaccc ctgagatacc tggtggagtt gcacctacac cacaatcctt ggaactgtga
     2521 ttgtgatatt ctgtggctgg cttggtggct tcgggagtat atcccaacca attccacctg
     2581 ctgtggccgc tgtcatgctc ccatgcacat gcgaggccgc tacctggtag aggtggacca
     2641 ggctgccttt cagtgctctg cccctttcat catggatgca ccccgggacc tcaatatctc
     2701 tgaggatcgg atggcagaac tgaagtgtcg gactccccct atgtcctctg tgaagtggtt
     2761 gctgcccaat gggacagtgc tcagccacgc ctcccggcat ccccggatct ctgttctcaa
     2821 tgatggcacc ttgaactttt ctcgtgtgct gctcatagac actggagtat acacatgcat
     2881 ggtgaccaat gtggcaggca actccaatgc ctcggcctac ctcaatgtga gctcggccga
     2941 gctcaacacc cccaacttca gcttcttcac cactgtaaca gtggaaacca cagagatatc
     3001 acctgaggac ataacacgga agtacaagcc tgttcccacc acatccactg gttaccagcc
     3061 ggcatatacc acctctacca cggtgctcat tcagaccacc cgtgtgccca agcaggtgcc
     3121 agtaccctcc acagatacca ctgacaagat gcagaccagc ctggatgaag tcatgaagac
     3181 caccaagact agtggatctt ccggaatcat cccagttgag gaggagaacc cggacttctg
     3241 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     3301 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     3361 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     3421 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     3481 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     3541 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     3601 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     3661 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     3721 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     3781 caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac
     3841 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     3901 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     3961 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     4021 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     4081 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     4141 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     4201 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     4261 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     4321 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     4381 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     4441 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     4501 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     4561 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     4621 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gtcatcatca
     4681 ccatcaccat taatctagag atcttattaa agcagaactt gtttattgca gcttataatg
     4741 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4801 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagcta
     4861 gactcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4921 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4981 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5041 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5101 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5161 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5221 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5281 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5341 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5401 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5461 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5521 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5581 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5641 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5701 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5761 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5821 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5881 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5941 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6001 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6061 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6121 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6181 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6241 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6301 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6361 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6421 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6481 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6541 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6601 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6661 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6721 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6781 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6841 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6901 cacatgcagc tcccggagga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6961 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     7021 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     7081 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7141 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     7201 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     7261 cgggagcttg tatatccatt ttcgtcccgg agacggtcac agcttgtctg taagcggatg
     7321 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     7381 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     7441 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     7501 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7561 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     7621 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     7681 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     7741 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     7801 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     7861 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     7921 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     7981 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     8041 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
