LOCUS       sequence-131136-        7799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Islr 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 7799)
  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 7799)
  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: Islr-AP-His
FEATURES             Location/Qualifiers
     source          1..7799
                     /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             4379..4396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4422..4543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4459..4478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4513..4532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4581..4598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4752..5340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4832..4851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5511..6302,6303..6371))
                     /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     6134..6153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6372..6476)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6544..6562
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6600..6622)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6624..6644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6629..6959
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6810..6945
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6872..6891
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7096..7115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7182..7637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7319..7340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7531..7550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7763..7785
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7777..7794
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7778..7794
                     /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 tgcgggcact
     1621 gtgcctgctt tgttgggctg ttctcctgaa cctggtgcga gcttgtccag agccctgtga
     1681 ctgtggggag aagtatggct tccagattgc agactgtgcc tatcgtgacc tagagggtgt
     1741 gccaccgggc ttcccggcca atgtgaccac actgagcctg tcagccaaca ggctgccagg
     1801 cctgccggag ggagccttca gggaagtgcc cctattgcag tcgctgtggc tggcacacaa
     1861 tgagatccgc tcggtggcta ttggtgctct ggcccctctg agccatctca agagtctgga
     1921 cctcagccac aacctcctct ctgagtttgc ctggagcgac ctgcacaacc tcagcgctct
     1981 gcagctgctc aagatggaca gcaacgagtt agccttcatc cctcgggacg cctttagcag
     2041 cctcagcgcc ctgcgttccc tgcagctcaa ccacaaccgc ctgcacgcgt tggccgaggg
     2101 caccttcgca ccgctcaccg cgctgtccca cttgcagatc aatgacaacc ctttcgactg
     2161 cacctgtggc atcgtgtggt tcaagacatg ggccctggcc tcagcggtgt ctattccaga
     2221 acaggacaac attgcctgta ctacacccca cgtcctgaag ggtatcccac taggccgcct
     2281 gccacccctg ccctgctcag ctccctcagt gcaactaagc taccagccca gccaagatgg
     2341 agcagagcta cggcctggct tcgtgctggc actccactgt gacgtggatg gacagccagt
     2401 cccccagctc cattggcaca ttcacacccc gggcggcacg gtggagatcg ccagtcctaa
     2461 tgtaggcact gatggacgtg ccctgcctgg tgcccttgca accagtgggc agccacgctt
     2521 ccaggccttt gccaatggca gcctgcttat ccctgacttt ggcaagctgg aggagggcac
     2581 ctatagctgc ctggccacca atgagctagg cagtgccgaa agctctgtaa atgtggcatt
     2641 ggccacccca ggtgaggggg gagaggatgc tgtggggcac aagttccatg gcaaagcagt
     2701 ggagggcaag ggctgctata cggttgacaa cgaggtacag ccatccggac cggaagacaa
     2761 cgtggttatc atttacctca gccgtgctgg gcccccagaa gctgcaatag cagcagacgg
     2821 gaggcctgcg cagcagttct ctggcatact tctgctaggc caaagcctgc ttgttctctc
     2881 ctttttctac ggcgcgcctg gatcttccgg aatcatccca gttgaggagg agaacccgga
     2941 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     3001 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     3061 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     3121 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     3181 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     3241 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     3301 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     3361 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     3421 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     3481 catctccaac atggacattg acgtgatcct aggtgggggc cgaaagtaca tgtttcgcat
     3541 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     3601 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtacg tgtggaaccg
     3661 cactgagctc atgcaggctt ccctggaccc gtctgtggcc catctcatgg gtctctttga
     3721 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     3781 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     3841 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     3901 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     3961 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     4021 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     4081 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     4141 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     4201 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     4261 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     4321 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtca
     4381 tcatcaccat caccattaat ctagagatct tattaaagca gaacttgttt attgcagctt
     4441 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4501 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggtcgactc
     4561 tagctagact cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4621 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4681 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4741 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4801 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4861 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4921 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4981 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5041 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5101 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5161 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5221 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5281 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5341 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5401 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5461 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5521 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5581 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5641 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5701 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5761 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5821 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5881 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5941 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6001 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6061 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6121 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6181 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6241 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6301 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6361 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6421 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6481 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6541 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     6601 ctctgacaca tgcagctccc ggaggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6661 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6781 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6841 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     6901 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     6961 agctcccggg agcttgtata tccattttcg tcccggagac ggtcacagct tgtctgtaag
     7021 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7081 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7141 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     7201 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     7261 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     7321 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     7381 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     7441 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     7501 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     7561 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     7621 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     7681 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     7741 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatt
//
