LOCUS       sequence-135793-        8153 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the FLRT1 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 8153)
  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 8153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: Flrt1-AP-His
FEATURES             Location/Qualifiers
     source          1..8153
                     /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"
     regulatory      1688..1697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4733..4750
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4776..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4813..4832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4867..4886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4935..4952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5106..5694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5186..5205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5865..6656,6657..6725))
                     /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     6488..6507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6726..6830)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6898..6916
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6954..6976)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6978..6998)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6983..7313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7164..7299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7226..7245
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7450..7469
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7536..7991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7673..7694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7885..7904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8117..8139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8131..8148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8132..8148
                     /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 tggtggtggc
     1621 acactctgct gccaccgcta ccaccacacc tgctgccacg gtcacagcca ctgtcgtgat
     1681 gaccacagcc accatggacc tgcgggactg gctgtttctc tgttatgggc tcattgcctt
     1741 cctcacggag gtcatcgata gcaccacctg cccatcagtg tgccgctgtg acaatggctt
     1801 catctactgc aatgaccggg gactcacatc catcccctca gacatccccg acgatgccac
     1861 caccctctac ctacagaaca accagatcaa caatgcgggc atccctcagg acctcaagac
     1921 caaggtcaag gtgcaggtca tctacctgta tgagaatgac ctggacgagt tccctatcaa
     1981 tctcccccgc tccctgagag agctgcatct gcaggacaac aatgtgcgca ccatcgccag
     2041 ggactctttg gctcgcatcc cgctgctgga gaagctgcac ctggacgaca actccgtttc
     2101 caccgtgagc atcgaggagg atgcttttgc tgacagcaag cagctcaagc tgcttttcct
     2161 gagccggaac cacctgagca gcatcccgtc agggctgccc cacacgctgg aggaactgag
     2221 gctagatgac aaccgcatct ctaccatccc cctgcacgca ttcaagggtc tcaacagcct
     2281 gcggcgcctg gttctggatg gcaacctgct ggccaaccag cgcattgccg acgacacctt
     2341 cagccggctg cagaacctca cggagctgtc gctagtacgg aactcgctgg ctgccccgcc
     2401 ccttaatctg cccagcgccc acctgcagaa gctctacctg caggacaatg ccatcagtca
     2461 cattccctac aacaccctgg ccaagatgcg ggagctggag aggttggacc tgtccaacaa
     2521 caacctcacc acgctgcctc ggggcctgtt tgatgacctg gggaacctgg cacagctgct
     2581 tctcaggaac aacccctggt tctgtggctg taacctcatg tggctgaggg actgggtgag
     2641 ggcacgggct gcagtggtca atgtgcgggg cctcatgtgc cagggccctg agaaggtccg
     2701 gggcatggcc atcaaagaca tcaccagcga gatggatgag tgctttgagg cggggtcaca
     2761 gggcggtgct gctaatgcag ccgccaagac cacagtcagc aaccatgcct ctgccaccac
     2821 accccagggc tctctgttta ccctcaaggc caagaggcca ggactgcgcc tcccagactc
     2881 caacattgac taccccatgg ccactggcga tggcgccaag acattggtca tccaggtgaa
     2941 gccactgacg gcagactcta tccgaatcac atggaaggcc atgctacccg cctcttcttt
     3001 ccggctcagt tggctacgtc tgggccatag cccggctgtg ggctctataa cagagaccct
     3061 ggtgcagggg gacaagacag agtacttgct gacagcactg gagcccaagt ccacctacat
     3121 catctgtatg gtcaccatgg agactggcaa cacctacgtg gccgatgaga cacctgtgtg
     3181 tgccaaggca gagacagcag atagctatgg ccctaccacc acgctcaacc aggaacagaa
     3241 tgctggcact agtggatctt ccggaatcat cccagttgag gaggagaacc cggacttctg
     3301 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     3361 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     3421 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     3481 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     3541 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     3601 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     3661 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     3721 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     3781 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     3841 caacatggac attgacgtga tcctaggtgg gggccgaaag tacatgtttc gcatgggaac
     3901 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     3961 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     4021 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     4081 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     4141 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     4201 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     4261 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     4321 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     4381 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     4441 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     4501 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     4561 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     4621 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     4681 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gtcatcatca
     4741 ccatcaccat taatctagag atcttattaa agcagaactt gtttattgca gcttataatg
     4801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagcta
     4921 gactcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5521 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5761 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5821 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5881 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5941 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6001 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6061 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6121 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6181 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6241 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6301 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6361 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6421 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6481 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6541 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6601 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6661 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6721 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6781 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6841 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6901 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6961 cacatgcagc tcccggagga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     7021 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     7081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     7141 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7201 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     7261 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     7321 cgggagcttg tatatccatt ttcgtcccgg agacggtcac agcttgtctg taagcggatg
     7381 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     7441 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     7501 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     7561 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7621 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     7681 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     7741 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     7801 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     7861 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     7921 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     7981 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8041 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     8101 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
