LOCUS       sequence-131179-        7403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G1, isoform j 
            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 7403)
  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 7403)
  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.j-AP-His
FEATURES             Location/Qualifiers
     source          1..7403
                     /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             3983..4000
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4026..4147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4063..4082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4117..4136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4185..4202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4356..4944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4436..4455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5115..5906,5907..5975))
                     /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     5738..5757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5976..6080)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6148..6166
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6204..6226)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6228..6248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6233..6563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6414..6549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6476..6495
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6700..6719
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6786..7241)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6923..6944)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7135..7154
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7367..7389
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7381..7398
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7382..7398
                     /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 aggaaggact agtggatctt ccggaatcat
     2521 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     2581 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     2641 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     2701 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     2761 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2821 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2881 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     2941 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     3001 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     3061 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     3121 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     3181 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     3241 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     3301 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     3361 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     3421 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     3481 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     3541 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     3601 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     3661 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     3721 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3781 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3841 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3901 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     3961 caccgacgcc gcgcacccgg gtcatcatca ccatcaccat taatctagag atcttattaa
     4021 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4081 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4141 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg
     4201 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4261 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4321 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4381 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4441 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4501 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4561 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4621 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4681 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4741 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4801 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4861 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4921 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4981 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5041 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5101 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5161 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5221 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5281 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5341 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5401 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5461 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5521 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5581 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5641 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5701 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5761 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5821 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5881 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5941 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6001 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6061 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6121 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6181 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca
     6241 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     6301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6481 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6541 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg
     6601 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6661 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     6721 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6781 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     6841 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     6901 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     6961 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7021 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7081 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7141 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7201 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     7261 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7321 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7381 tgtaaaacga cggccagtga att
//
