LOCUS       sequence-131166-        8402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 4 protein, 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 8402)
  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 8402)
  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: Ntn4-AP-His
FEATURES             Location/Qualifiers
     source          1..8402
                     /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)"
     regulatory      1604..1613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4982..4999
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5025..5146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5062..5081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5116..5135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5184..5201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5355..5943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5435..5454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6114..6905,6906..6974))
                     /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     6737..6756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6975..7079)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7147..7165
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7203..7225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7227..7247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7232..7562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7413..7548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7475..7494
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7699..7718
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7785..8240)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7922..7943)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8134..8153
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8366..8388
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8380..8397
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8381..8397
                     /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 tggggagctg
     1621 cgcacggctg ctgctgctct ggggctgctc cgcggtggcc gcaggcttga atggagtagc
     1681 cggagcgaac tcccgctgtg agaaggcatg caaccctcgc atgggaaact tggctttggg
     1741 aagaaagctc cgggcagaca ctatgtgtgg ccagaacgcc accgaactct tctgcttcta
     1801 cagtgagaat gctgacctca cttgccggca gcccaagtgt gataaatgca acgctgccca
     1861 ttctcaccta gctcacccac cctctgccat ggcagactca tccttcaggt ttccccggac
     1921 atggtggcag tctgcagagg atgtgcacag ggaaaagatt cagctagacc tggaagcaga
     1981 attctacttc actcacctaa ttatggtgtt caagtctccc aggcctgcag ccatggtgct
     2041 ggaccggtcc caggactttg ggaagacctg gaagccttac aagtactttg caacaaactg
     2101 ctcggctact tttggcctgg aagatgatgt tgtcaagaag ggagctattt gcacgtctag
     2161 atactcaaat cctttcccgt gcaccggagg agaggttatt ttcagagccc tgtcaccacc
     2221 atacgacata gaaaaccctt acagtgccaa agtgcaggag cagctgaaga tcaccaacct
     2281 ccgagtgcgg ctgctcaagc gacagtcctg cccttgtcag ataaacgacc tgaacgcaaa
     2341 acctcaccat tttatgcact acgcagtcta tgacttcatc gtcaagggca gctgcttctg
     2401 caacggccac gctgaccagt gcttacctgt ggagggcttc agacccatca aggccccggg
     2461 agcgttccac gtggtccacg ggaggtgtat gtgtaagcac aacacagcag gcagccactg
     2521 ccagcactgt gcaccattgt acaatgaccg gccctgggag gcagcggacg gcagaacagg
     2581 ggctcctaac gaatgcagaa cttgcaagtg caatgggcac gcggacacct gtcacttcga
     2641 cgtcaacgtg tgggaggcgt cggggaaccg cagcggcggt gtctgcaaca actgtcagca
     2701 caacactgag ggtcagcact gtcagcgctg taagcccggt ttctaccgcg acctcagaag
     2761 acccttctcc gcccctgacg cttgcaaagc gtgttcctgc cacccggttg gatcagcgat
     2821 ccttcctttc agctcagtga ccttctgcga ccccagcaat ggtgactgcc cctgcaagcc
     2881 tggggtggcg gggccacatt gtgacagatg catggtggga tactggggtt ttggagacta
     2941 cggctgcaga ccttgcgatt gtgcggggag ctgcgacccg ctcacgggag actgcatcag
     3001 cagtaacgct gatgtagact ggtaccacga agtccccacc tttcactcga tgcacaataa
     3061 gagtgagccc agctgggaat gggaggatga gcaaggattt tctgccctcc gacactcagg
     3121 taaatgtgaa tgtaaggaac aggtgttagg aaaccccaaa gccttctgtg gaatgaagta
     3181 ttcatatgtg ttaaaaatca agatcttatc agcccatgac aaaggctccc atgccgaagt
     3241 caatgtgaag attaagaaag tcttaaagtc caccaaactg aagatcttac gaggcaagag
     3301 aacgctatac ccagagtcct ggactaacag aggctgcacc tgtccaatcc tcaatccagg
     3361 attggagtac ctggtcgccg gccacgagga cgtaagaacg ggcaaattaa ttgtgaatat
     3421 gaaaagcttt gtccagcact ggaaaccagc tcttggcaga agagtcatgc acatcttaaa
     3481 aagagactgc gtgggcgcgc ctggatcttc cggaatcatc ccagttgagg aggagaaccc
     3541 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     3601 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     3661 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     3721 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     3781 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     3841 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     3901 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     3961 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     4021 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     4081 gctcatctcc aacatggaca ttgacgtgat cctaggtggg ggccgaaagt acatgtttcg
     4141 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     4201 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt acgtgtggaa
     4261 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg gcccatctca tgggtctctt
     4321 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     4381 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     4441 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     4501 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     4561 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     4621 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     4681 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     4741 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     4801 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     4861 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     4921 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     4981 tcatcatcac catcaccatt aatctagaga tcttattaaa gcagaacttg tttattgcag
     5041 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5101 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggtcga
     5161 ctctagctag actcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5761 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7081 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7141 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     7201 aacctctgac acatgcagct cccggaggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     7261 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     7321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     7381 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7441 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7501 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     7561 aaaagctccc gggagcttgt atatccattt tcgtcccgga gacggtcaca gcttgtctgt
     7621 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     7681 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     7741 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     7801 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     7861 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     7921 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     7981 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     8041 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     8101 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     8161 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     8221 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     8281 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     8341 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     8401 tt
//
