LOCUS       sequence-131134-        8132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the FLRT2 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 8132)
  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 8132)
  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: Flrt2-AP-His
FEATURES             Location/Qualifiers
     source          1..8132
                     /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             4712..4729
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4755..4876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4792..4811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4846..4865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4914..4931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5085..5673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5165..5184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5844..6635,6636..6704))
                     /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     6467..6486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6705..6809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6877..6895
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6933..6955)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6957..6977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6962..7292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7143..7278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7205..7224
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7429..7448
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7515..7970)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7652..7673)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7864..7883
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8096..8118
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8110..8127
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8111..8127
                     /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 tgggcctaca
     1621 gactacaaag tggcccggcc gtggggcttt catcctcaaa ttttggctca tcatttccct
     1681 gggactctac ttacaagtgt caaaactcct ggcctgccct agtgtatgcc gctgtgacag
     1741 gaactttgtc tactgtaacg agagaagctt gacctcagtg cctcttggga tcccggaggg
     1801 cgtaaccgta ctctacctcc acaacaacca aattaataat gctggctttc ctgcagagct
     1861 gcacaatgtc cagtcagtgc acactgtcta cctttatggc aaccagttgg atgagttccc
     1921 catgaacctt cccaagaatg tcagagttct ccatctgcag gaaaacaaca ttcagaccat
     1981 ctcgcgggct gctctcgctc agctcctcaa gctggaagaa ctccacctgg acgacaactc
     2041 catatctaca gtgggagtag aagacggagc gttccgggag gcgattagcc tcaaactgtt
     2101 gtttttatcg aagaatcacc tgagtagcgt gcctgttggg cttcctgtag acttgcaaga
     2161 gctgagagtg gatgaaaacc gaattgccgt catatcagac atggcctttc agaacctcac
     2221 aagcttggag cgcctgatcg tggatgggaa tcttctgacc aacaagggca ttgctgaggg
     2281 taccttcagt catctcacca agctcaagga attttctata gtccggaact cgctctccca
     2341 cccacctcct gatctcccag gtacgcatct gatcaggctc tacttgcagg ataaccagat
     2401 aaaccatatc ccattgacag ccttcgcaaa cctccgtaag ctggaaaggc tagatatatc
     2461 caacaaccag ctacgaatgt taactcaagg agtcttcgat catctctcca acctgaaaca
     2521 actcactgcg cggaataacc cttggttttg tgactgcagc attaaatggg tcacagaatg
     2581 gctcaagtat atcccttctt ctctcaacgt gcgtggtttc atgtgccaag gtcctgagca
     2641 agtccgcggg atggctgtca gggagttgaa tatgaatctt ttgtcttgtc ccaccacgac
     2701 tcctggccta cctgtcttta ccccagctcc aagtaccgtt tctccaacaa ctcagtctcc
     2761 tactctctct gttccaagcc ccagcagagg ctctgtgcct ccagctccta ccccatcgaa
     2821 acttcccacc atccctgatt gggatggcag agaaagagtg accccaccta tttctgaaag
     2881 gatccaactt tccatccact ttgttaatga tacttcgatt caagtcagct ggctgtctct
     2941 ctttactgtg atggcctaca aactgacatg ggtgaaaatg ggccacagtc tcgtaggggg
     3001 catcgttcag gaacgaattg tcagtggtga gaagcaacac ctgagcttgg ttaatttaga
     3061 gcccagatcc acgtatagga tttgtttagt gccgctggat gcgttcaact accgcactgt
     3121 ggaagatacc atctgttcgg aggctaccac ccatgcctct tatttgaaca acggcagcaa
     3181 cactgcttct agccatgagc agacgacttc ccacagtatg ggctccacta gtggatcttc
     3241 cggaatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     3301 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     3361 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     3421 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     3481 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     3541 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     3601 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     3661 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     3721 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     3781 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     3841 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     3901 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     3961 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     4021 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     4081 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     4141 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     4201 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     4261 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     4321 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     4381 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     4441 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     4501 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     4561 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     4621 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     4681 cgccggcacc accgacgccg cgcacccggg tcatcatcac catcaccatt aatctagaga
     4741 tcttattaaa gcagaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4801 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4861 tcatcaatgt atcttatcat gtctggtcga ctctagctag actcttccgc ttcctcgctc
     4921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5761 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5821 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5881 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5941 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6001 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6061 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6121 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6181 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6241 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6301 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6361 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6421 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6481 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6541 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6601 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6661 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6721 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6781 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6841 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6901 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggaggaa
     6961 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     7021 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     7081 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     7141 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     7201 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     7261 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     7321 tcgtcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     7381 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     7441 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7501 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     7561 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7621 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     7681 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     7741 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     7801 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     7861 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     7921 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     7981 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     8041 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     8101 tcacgacgtt gtaaaacgac ggccagtgaa tt
//
