LOCUS       sequence-131180-        7858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G2, 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 7858)
  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 7858)
  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.a-AP-His
FEATURES             Location/Qualifiers
     source          1..7858
                     /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             4438..4455
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4481..4602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4518..4537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4572..4591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4640..4657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4811..5399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4891..4910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5570..6361,6362..6430))
                     /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     6193..6212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6431..6535)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6603..6621
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6659..6681)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6683..6703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6688..7018
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6869..7004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6931..6950
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7155..7174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7241..7696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7378..7399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7590..7609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7822..7844
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7836..7853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7837..7853
                     /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 ctttggcaac tgcgagtgct acggccactc caaccgttgc
     2701 agctacattg acttcctgaa cgtggtgacc tgcgtcagct gtaaacacaa cactcgaggc
     2761 caacactgtc agcactgccg cctgggctac tatcgcaatg gctccgcaga gctggatgat
     2821 gagaacgtct gcattgaatg taactgtaac cagatcggct ctgtgcacga tcgatgcaat
     2881 gagacaggct tctgcgagtg cagggagggc gcagtggggc ccaagtgcga cgactgcctt
     2941 cctacacact acactagtgg atcttccgga atcatcccag ttgaggagga gaacccggac
     3001 ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca gcctgcacag
     3061 acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc tacggtgaca
     3121 gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat acccctggcc
     3181 atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa acatgtgcca
     3241 gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt ccagaccatt
     3301 ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc
     3361 tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac caccacacga
     3421 gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa ctggtactcg
     3481 gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc tacgcagctc
     3541 atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg
     3601 ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag gctggacggg
     3661 aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt gtggaaccgc
     3721 actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg tctctttgag
     3781 cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc cctgatggag
     3841 atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt cctcttcgtg
     3901 gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc actgactgag
     3961 acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga ggaggacacg
     4021 ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta ccccctgcga
     4081 gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc ctacacggtc
     4141 ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc
     4201 gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct ggacgaagag
     4261 acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac
     4321 ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg cctggagccc
     4381 tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca cccgggtcat
     4441 catcaccatc accattaatc tagagatctt attaaagcag aacttgttta ttgcagctta
     4501 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4561 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggtcgactct
     4621 agctagactc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5461 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5521 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5641 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5701 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5761 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5821 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5881 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5941 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6001 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6061 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6541 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6601 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6661 tctgacacat gcagctcccg gaggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     6721 tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     6781 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6841 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6901 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     6961 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     7021 gctcccggga gcttgtatat ccattttcgt cccggagacg gtcacagctt gtctgtaagc
     7081 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7141 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     7261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     7381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     7501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     7621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7681 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     7741 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     7801 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaatt
//
