LOCUS       sequence-135571-        8288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the FLRT3 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 8288)
  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 8288)
  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: Islr2.b-AP-His
FEATURES             Location/Qualifiers
     source          1..8288
                     /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             4868..4885
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4911..5032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4948..4967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5002..5021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5070..5087)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5241..5829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5321..5340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6000..6791,6792..6860))
                     /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     6623..6642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6861..6965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7033..7051
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7089..7111)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7113..7133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7118..7448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7299..7434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7361..7380
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7585..7604
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7671..8126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7808..7829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8020..8039
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8252..8274
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8266..8283
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8267..8283
                     /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 tggggccctt
     1621 tggagccctg tgtttggcct gggctttgct aggagtggtc agagcgtgtc ccgagccttg
     1681 cgcctgtgtt gacaagtacg cccaccagtt tgcagactgt gcctacaagg agctgcgcga
     1741 ggtcccagaa ggactgccag ccaacgtgac cacgcttagt ctgtctgcca acaagattac
     1801 ggtactaagg cggggggcct tcgtcaacgt cacgcaggtc acttcgctgt ggctggctca
     1861 cagtgaggta cgcacggtag agtcaggggc attggcagtg ctgagtcagc tcaagaacct
     1921 cgacctaagc cacaacctta tatccaactt cccttggagc gaccttcgta acttgagcgc
     1981 gctgcagctg ctgaaaatga accacaaccg tctgggatcg ctgccccggg atgcactcgg
     2041 cgcgctcccg gacctgcgct ctctgcgcat caacaacaac cggctgcgta ccctggagcc
     2101 cggcacgttc gacgcactaa gcgcgctgtc tcacctgcaa ctctatcaca accccttcca
     2161 ctgcagctgt ggtctcgtgt ggctgcaggc ctgggcagcg agcacccggg tctccttacc
     2221 ggagcccgat tctatagcgt gcgcctcgcc ccctgagctg cagggcgtgc ccgtgcaccg
     2281 cctgcccgcc ctgccctgcg caccccccag cgtgcgtctg agcgcagagc cgccgcctga
     2341 ggcacctggc acccctctgc gcgcaggctt ggctttcatg ttacattgcg tcgccgaagg
     2401 ccaccccaca ccccgcctgc aatggcaact tcagatcccg ggtggcactg tagtcttagt
     2461 gccaccggtt ctcagcaagg aagaagatgg aggagataag gtggaggatg gagagggtga
     2521 cggagatgag gacctgccta cgcagactga agcgccaacc ccaactccag cacctgcttg
     2581 gccagcgcct ccagccaccc cgcgcttctt ggccctcgca aatggctctc tgttggtgcc
     2641 ccttctgagt gccaaggagg caggcatcta cacttgtcgt gcacacaatg agctgggcac
     2701 caactcaacg tccttacggg tgacggtggc tgcagcgggg ccgccaaaac acgctcctgg
     2761 aacaggggaa gaacctgatg cgcaggtccc gacctctgag cgcaaggcca ccactaaggg
     2821 ccgtagcaac agcgtcctgc ccttcaagcc tgaaggcaaa accaaaggcc aaggtcttgc
     2881 ccgagtcagc gtgcttgggg aaatcgaggc agagctggag gagacagatg aaggagagca
     2941 gatggaaggt cagatccctg cagatccgat gggagagaag cactgtggcc atggggaccc
     3001 ctctcgctat gtgtctaacc atgcattcaa ccagagctca gatctcaagc cgcacgtttt
     3061 tgagttgggc gtcatcgcgc tggatgtagc agagcgtgag gctcgggtgc agctgacgcc
     3121 ccttgctgcg cgctggggcc ctgggccaga tggtgctagc ggagcgcggc ggccggggag
     3181 gcggccactg cgcctactct atctgtgtcc tgcggggggt ggcacggcag ttcagtggtc
     3241 acgagtggaa gagggggtca atgcctactg gtttcgcggc ctgcggcctg gcaccaacta
     3301 ctccgtatgc ctggcactgg cgggcgaggc gtgtcacgtg caggtggtgt tttctaccaa
     3361 gaaagaactg ccgtccctgg gcgcgcctgg atcttccgga atcatcccag ttgaggagga
     3421 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     3481 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     3541 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     3601 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     3661 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     3721 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     3781 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     3841 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     3901 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     3961 tacgcagctc atctccaaca tggacattga cgtgatccta ggtgggggcc gaaagtacat
     4021 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     4081 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     4141 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     4201 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     4261 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     4321 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     4381 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     4441 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     4501 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     4561 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     4621 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     4681 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     4741 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     4801 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     4861 cccgggtcat catcaccatc accattaatc tagagatctt attaaagcag aacttgttta
     4921 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4981 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5041 ggtcgactct agctagactc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5101 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5161 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5221 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5401 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5641 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5701 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5761 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5821 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5881 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5941 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6001 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6061 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6121 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6181 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6241 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6301 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6361 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6421 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6481 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6541 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6601 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6661 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6721 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6781 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6841 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6961 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7021 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     7081 ggtgaaaacc tctgacacat gcagctcccg gaggaatgtg tgtcagttag ggtgtggaaa
     7141 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     7201 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     7261 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     7321 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     7381 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     7441 tttgcaaaaa gctcccggga gcttgtatat ccattttcgt cccggagacg gtcacagctt
     7501 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     7561 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7621 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     7681 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     7741 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     7801 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     7861 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     7921 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     7981 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     8041 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     8101 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     8161 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     8221 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     8281 agtgaatt
//
