LOCUS       sequence-131172-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G1, isoform c 
            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 7712)
  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 7712)
  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.c-AP-His
FEATURES             Location/Qualifiers
     source          1..7712
                     /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             4292..4309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4335..4456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4372..4391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4426..4445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4494..4511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4665..5253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4745..4764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5424..6215,6216..6284))
                     /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     6047..6066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6285..6389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6457..6475
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6513..6535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6537..6557)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6542..6872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6723..6858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6785..6804
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7009..7028
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7095..7550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7232..7253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7444..7463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7676..7698
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7690..7707
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7691..7707
                     /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 tcggtactaa
     2701 tgtctgcgac aatgagctcc tgcactgcca gaatggaggg acctgccaga acaatgtgcg
     2761 ctgcgcgtgc ccagacgcct acaccggcat cctctgtgag aagctaacta gtggatcttc
     2821 cggaatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     2881 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     2941 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     3001 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     3061 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     3121 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     3181 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     3241 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     3301 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     3361 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     3421 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     3481 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     3541 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     3601 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     3661 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     3721 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     3781 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     3841 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     3901 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     3961 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     4021 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     4081 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     4141 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     4201 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     4261 cgccggcacc accgacgccg cgcacccggg tcatcatcac catcaccatt aatctagaga
     4321 tcttattaaa gcagaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4381 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4441 tcatcaatgt atcttatcat gtctggtcga ctctagctag actcttccgc ttcctcgctc
     4501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6421 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6481 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggaggaa
     6541 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     6601 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6661 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6721 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6781 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6841 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     6901 tcgtcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6961 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     7021 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7081 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     7141 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7201 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     7261 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     7321 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     7381 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     7441 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     7501 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     7561 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7621 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7681 tcacgacgtt gtaaaacgac ggccagtgaa tt
//
