LOCUS       sequence-131181-        8035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G2, isoform b 
            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 8035)
  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 8035)
  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: Ntng2.b-AP-His
FEATURES             Location/Qualifiers
     source          1..8035
                     /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             4615..4632
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4658..4779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4695..4714)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4749..4768
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4817..4834)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4988..5576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5068..5087)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5747..6538,6539..6607))
                     /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     6370..6389
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6608..6712)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6780..6798
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6836..6858)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6860..6880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6865..7195
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7046..7181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7108..7127
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7332..7351
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7418..7873)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7555..7576)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7767..7786
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7999..8021
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8013..8030
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8014..8030
                     /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 caaacgccac catgctgcgc
     1621 ctgctggctc tcttcctgca ctgcctcccg ctggtctctg gggactatga catctgcaaa
     1681 tcctgggtga ccacagatga gggccccacc tgggaattct atgcctgcca gcccaaggtg
     1741 atgcgcctga aggattatgt caaggtcaaa gtggagccct caggcatcac gtgcggagac
     1801 ccccctgaaa ggttctgttc ccatgagaac ccctacctgt gcagtaatga gtgtgatgcc
     1861 tccaatcccg acctggccca cccgccccgg cttatgtttg acagggaaga tgagggactg
     1921 gctacctact ggcaaagcgt cacgtggagt cgctacccca gtccactaga ggccaacatc
     1981 accctctcat ggaacaagag cgtggagttg acagacgacg tggtggtgac ttttgagtat
     2041 ggccggccca cggtcatggt cctcgagaag tccctggaca atggccgcac ctggcagccc
     2101 taccagttct atgcagagga ctgcatggag gccttcggca tgtctgcccg acgtgcccga
     2161 gacatgtcac cctccagcgc ccaccgggtg ctctgcaccg aggagtactc acgctgggca
     2221 gggtccaaga aagagaagca tgtgcgcttt gaggtaaggg accgctttgc catctttgcc
     2281 ggccctgacc tgcgtaacat ggacaacctg tacacgagga tggagagcgc caagggcctc
     2341 aaggagttct tcaccttcac tgacctgcgc atgcgcctgc tgcgtcctgc gctgggtggc
     2401 acctacgtgc agcgagagaa cctctacaag tacttctatg ccatctccaa tatcgaagtc
     2461 attggcaggt gtaagtgcaa cctgcatgcc aacctgtgca cagtgcgaga gggcagcctg
     2521 cagtgtgagt gtgaacacaa caccacgggc cccgactgtg gcaggtgcaa gaagaacttc
     2581 cgcacacgcg cctggcgagc tggctcctac ctgccgctgc cccacggctc tcccaatgcc
     2641 tgtgcggccg cgggctccgc ctttggcagt cagaccaagc cacctactat ggcccccctt
     2701 ggggacagct ccttctggcc ccaggtgtcc tccagtgcag aagctgtagc tatctctgtc
     2761 gctgtccctt cccaagccaa ggactctacg cttttcgagc tcaagcccag atctcctcag
     2821 gtgataccca ttgaagaatt tcaagactgc gagtgctacg gccactccaa ccgttgcagc
     2881 tacattgact tcctgaacgt ggtgacctgc gtcagctgta aacacaacac tcgaggccaa
     2941 cactgtcagc actgccgcct gggctactat cgcaatggct ccgcagagct ggatgatgag
     3001 aacgtctgca ttgaatgtaa ctgtaaccag atcggctctg tgcacgatcg atgcaatgag
     3061 acaggcttct gcgagtgcag ggagggcgca gtggggccca agtgcgacga ctgccttcct
     3121 acacactaca ctagtggatc ttccggaatc atcccagttg aggaggagaa cccggacttc
     3181 tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc tgcacagaca
     3241 gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac ggtgacagct
     3301 gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc cctggccatg
     3361 gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca tgtgccagac
     3421 agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca gaccattggc
     3481 ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga ggtcatctcc
     3541 gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac cacacgagtg
     3601 cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg gtactcggac
     3661 gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac gcagctcatc
     3721 tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt tcgcatggga
     3781 accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct ggacgggaag
     3841 aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact
     3901 gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct ctttgagcct
     3961 ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct gatggagatg
     4021 acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct cttcgtggag
     4081 ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact gactgagacg
     4141 atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga ggacacgctg
     4201 agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg
     4261 agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta cacggtcctc
     4321 ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga tgttaccgag
     4381 agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga cgaagagacc
     4441 cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc
     4501 gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct ggagccctac
     4561 accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc gggtcatcat
     4621 caccatcacc attaatctag agatcttatt aaagcagaac ttgtttattg cagcttataa
     4681 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4741 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggt cgactctagc
     4801 tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4861 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4921 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4981 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5041 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5101 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5161 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5221 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5281 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5341 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5401 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5461 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5521 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5581 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5641 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5701 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5761 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5821 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5881 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5941 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6001 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6061 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6121 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6181 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6241 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6301 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6361 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6421 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6481 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6541 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6601 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6661 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6721 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6781 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6841 gacacatgca gctcccggag gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     6901 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     6961 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7021 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     7081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     7141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     7201 cccgggagct tgtatatcca ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga
     7261 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     7321 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     7381 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     7441 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7501 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7561 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7621 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7681 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7741 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7801 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7861 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     7921 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     7981 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaatt
//
