LOCUS       sequence-131176-        7880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G1, isoform g 
            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 7880)
  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 7880)
  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.g-AP-His
FEATURES             Location/Qualifiers
     source          1..7880
                     /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             4460..4477
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4503..4624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4540..4559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4594..4613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4662..4679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4833..5421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4913..4932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5592..6383,6384..6452))
                     /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     6215..6234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6453..6557)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6625..6643
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6681..6703)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6705..6725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6710..7040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6891..7026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6953..6972
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7177..7196
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7263..7718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7400..7421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7612..7631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7844..7866
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7858..7875
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7859..7875
                     /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 atagctaatg tctgcgacaa
     2881 tgagctcctg cactgccaga atggagggac ctgccagaac aatgtgcgct gcgcgtgccc
     2941 agacgcctac accggcatcc tctgtgagaa gctaactagt ggatcttccg gaatcatccc
     3001 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     3061 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     3121 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     3181 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     3241 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     3301 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     3361 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     3421 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     3481 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     3541 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     3601 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     3661 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     3721 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     3781 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     3841 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     3901 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3961 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     4021 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     4081 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     4141 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     4201 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     4261 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     4321 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     4381 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     4441 cgacgccgcg cacccgggtc atcatcacca tcaccattaa tctagagatc ttattaaagc
     4501 agaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4561 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4621 cttatcatgt ctggtcgact ctagctagac tcttccgctt cctcgctcac tgactcgctg
     4681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5461 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5521 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5581 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5641 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5701 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5761 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5821 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5881 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5941 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6001 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6061 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6121 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6181 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6241 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6301 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6361 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6421 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6481 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6541 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6601 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6661 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggaggaatg tgtgtcagtt
     6721 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     6781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6961 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7021 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc gtcccggaga
     7081 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     7141 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7201 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7261 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7321 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7381 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7441 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7501 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     7561 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     7621 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     7681 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc
     7741 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     7801 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     7861 aaaacgacgg ccagtgaatt
//
