LOCUS       sequence-131170-        8015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G1, isoform a 
            protein (truncated immediately upstream of propeptide cleavage and 
            GPI-attachement site), 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 8015)
  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 8015)
  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: Ntng1.a-AP-His
FEATURES             Location/Qualifiers
     source          1..8015
                     /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             4595..4612
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4638..4759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4675..4694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4729..4748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4797..4814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5727..6518,6519..6587))
                     /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     6350..6369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6588..6692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6760..6778
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6816..6838)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6840..6860)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6845..7175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7026..7161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7088..7107
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7312..7331
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7398..7853)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7535..7556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7747..7766
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7979..8001
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7993..8010
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7994..8010
                     /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 tgtatttgtc
     1621 aagattcctg tcgatccatg ccctgtgggt gacagtgtcc tctgtgatgc agccctacct
     1681 tttcgtgtgg ggacattatg atgtatgtaa gagcctgatt tacacagaag aaggcaaagt
     1741 ttgggattac acagcctgcc agccggaatc cacggacatg accaagtatc tgaaagtgaa
     1801 actggaccct ccggatatta cctgtggaga ccctccagag tccttctgtg caatgggcaa
     1861 cccttacatg tgcaataatg agtgtgatgc gagtacccct gaactggcac accctcctga
     1921 gctgatgttt gattttgaag gaagacatcc ctccacattt tggcagtctg ctacttggaa
     1981 ggagtacccc aaacctctcc aggttaacat cactctgtct tggagcaaaa ccattgaact
     2041 cacagacaac atagttatta cctttgaatc ggggcgtcca gaccaaatga tcctagagaa
     2101 atctctcgac tacggacgaa catggcagcc ctatcagtat tatgccacag actgcctcca
     2161 tgcattccac atggacccga aatccgtgaa ggatttatct cagcacacgg tcttggaaat
     2221 catttgcacg gaagagtact ccactgggta ctccacgaat agcaaaataa tccacttcga
     2281 gatcaaagac aggtttgcgt ttttcgctgg acctcggcta cgaaatatgg cttccctcta
     2341 tggacagctg gatacaacca agaaactcag agatttcttc actgtcacag acctgaggat
     2401 caggctgttg agacccgccg ttggggaaat atttgtagat gaactacatt tggcacgtta
     2461 cttttatgcg atctcagaca taaaggtgcg aggaaggtgc aagtgcaacc tgcatgccac
     2521 ttcgtgtttg tatgacaaca gcaaactgac atgtgaatgt gagcacaaca ctacaggtcc
     2581 cgactgtggg aaatgcaaga agaactacca gggccgacct tggagccccg gctcatacct
     2641 ccccatcccc aaaggcaccg caaacacctg tatccccagc atttccagta tcggtaactg
     2701 tgaatgcttc ggccactcca atcggtgcag ttatatcgat ctgctaaaca cagtcatttg
     2761 cgtgagctgt aaacacaaca ctagagggca gcactgtgag ttatgcaggc tgggctactt
     2821 cagaaatgct tctgcacagc tggacgatga gaatgtgtgc atagagtgtt attgtaaccc
     2881 tttgggctca atccatgatc gttgtaatgg ctcaggattt tgtgagtgta agactggaac
     2941 aacagggcct aaatgtgatg agtgtctgcc aggaaattcc tggtactacg gctgtcaacc
     3001 taatgtctgc gacaatgagc tcctgcactg ccagaatgga gggacctgcc agaacaatgt
     3061 gcgctgcgcg tgcccagacg cctacaccgg catcctctgt gagaagctaa ctagtggatc
     3121 ttccggaatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     3181 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     3241 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     3301 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     3361 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     3421 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     3481 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     3541 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     3601 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     3661 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     3721 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     3781 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     3841 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     3901 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     3961 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     4021 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     4081 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     4141 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     4201 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     4261 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     4321 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     4381 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     4441 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     4501 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     4561 ccccgccggc accaccgacg ccgcgcaccc gggtcatcat caccatcacc attaatctag
     4621 agatcttatt aaagcagaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4681 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4741 aactcatcaa tgtatcttat catgtctggt cgactctagc tagactcttc cgcttcctcg
     4801 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4861 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5641 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5701 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5761 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5821 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5881 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5941 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6001 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6061 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6121 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6181 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6241 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6301 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6361 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6421 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6481 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6541 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6601 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6661 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6721 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6781 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6841 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     6901 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     6961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     7021 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     7081 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     7141 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     7201 ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     7261 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     7321 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     7381 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7441 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     7501 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     7561 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     7621 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     7681 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     7741 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     7801 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     7861 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     7921 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     7981 cagtcacgac gttgtaaaac gacggccagt gaatt
//
