LOCUS       sequence-44831-e        7766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EXPRESs plasmid for Zebrafish Receptors encoding zebrafish unc5b 
            (netrin-1a) with rat CD4d3+4-bio-linker-His tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7766)
  AUTHORS   Sollner C, Wright GJ
  TITLE     A cell surface interaction network of neural leucine-rich repeat 
            receptors.
  JOURNAL   Genome Biol. 2009;10(9):R99. Epub 2009 Sep 18.
  PUBMED    19765300
REFERENCE   2  (bases 1 to 7766)
  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: "Genome 
            Biol."; date: "2009"; volume: "10(9)"; pages: "R99. Epub 2009 Sep 
            18"
COMMENT     Sequence Label: unc5bCD4d3+4-bio-linker-His
FEATURES             Location/Qualifiers
     source          1..7766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             644..661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    713..768
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(767..786)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3004..3022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3090..3194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3195..3263,3264..4055)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3413..3432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4226..4814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4715..4734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4968..4985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5102..5123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5138..5143,5144..5161,5162..5168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5176..5192
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5181..5203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5200..5216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5200..5216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5487..5866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5867..6070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6021..6041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6067..6091
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     6572..6591
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      6625..6634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtcgacctcc atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact
       61 caaccttgga gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc
      121 ttcccagtcc tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac
      181 tagcaacccc aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt
      241 ctcccttcag tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta
      301 tcaggaagtg aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg
      361 tgaggtgatg ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga
      421 ggccagggtc tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg
      481 gcaatgtcta ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc
      541 caaagggttg aattccggca gcctgcacca catcctggac gcccagaaga tgctgtggaa
      601 ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc atcaccacca
      661 ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg ctaataaagg
      721 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
      781 tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag aggatcgatc
      841 cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg cggggcagtg
      901 catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc cacatgtgac
      961 acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct tataaatgga
     1021 tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc agtctgtgga
     1081 ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca ccaatgctgg
     1141 gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa cgtcaatcag
     1201 aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa tagtgtttat
     1261 aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta gtatatacta
     1321 tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat gctatcgaat
     1381 tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga gctgtcacgg
     1441 ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt cacaagggca
     1501 gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt
     1561 ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg tatgtgaggt
     1621 gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg gggttcagtg
     1681 gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa taaatactag
     1741 tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt ggggacaagc
     1801 cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac ataatcctag
     1861 tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga caggtgaacc
     1921 atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca gcagcggact
     1981 ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa tggggcccat
     2041 aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca cgcgtcagcc
     2101 cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact tggctgattg
     2161 taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg
     2221 ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct gcgatctgga
     2281 ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt cctcatattc
     2341 acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata ctacccaaat
     2401 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     2461 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     2521 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     2581 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctaatagag
     2641 attagggtag tatatgctat cctaatttat atctgggtag catatactac ccaaatatct
     2701 ggatagcata tgctatccta atctatatct gggtagcata tgctatccta atctatatct
     2761 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     2821 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     2881 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     2941 gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa
     3001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3421 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3721 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     3781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4981 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5041 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5101 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5161 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5221 attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca
     5281 gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta
     5341 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
     5401 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     5461 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     5521 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     5581 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     5641 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     5701 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     5761 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     5821 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     5881 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     5941 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     6001 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     6061 aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg ctgtctgcga
     6121 gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg
     6181 gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg
     6241 catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta
     6301 ggctgagcac cgtggcgggc ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc
     6361 tgctgatgat gtaattaaag taggcggtct tgagacggcg gatggtcgag gtgaggtgtg
     6421 gcaggcttga gatccagctg ttggggtgag tactccctct caaaagcggg cattacttct
     6481 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgatctg
     6541 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     6601 caggtccaag tttaaactgc ggccgccacc atggtctcaa cgtgtataca ccgcgaccag
     6661 tctcccgcct ccctgctcgg tttacttcta ctcagctcaa ccttagtgat acatgcagat
     6721 ggcagcgatt acagtgaagt tctcccggac tcgtttccat cagctcctgc cgaaccgctg
     6781 ccagaattcc agagtgagcc cgaggatgca ttcatagtaa aaaacagacc cgtcaaactg
     6841 tcctgcaagg ccgcccctgc cacacagata tactttaaat gcaatggaga atgggtgaac
     6901 cagaatgacc atgtgaccaa ggaaagtctg gaccatatca caggtctggt agtgagagaa
     6961 gtagatatct ctgtctcccg gacgcaggta gaggagttgt ttgggctgga agattattgg
     7021 tgccagtgtg ttgcctggag ctcggcaggc accacaaaga gccgtcgggc ttacgtccgc
     7081 atcgcctact tgagaaagaa ctttgagcag gagccgcttg gcagggaggt gcgtctggag
     7141 caggaagtat tactgcagtg tcgtccacca gagggcagcc cgcctgctga ggtggactgg
     7201 ctaaagaacg aagagctcat tgacccggcg ctggattcta actttctaat taccatcgag
     7261 cacgacctga tcatcaaaca ggctcgactc tctgacactg ccaactacac ctgtgtggca
     7321 cgtaatgtgg tcgctaagag acgcagcagc actgccactc tcatcgtcta cgtaagtgga
     7381 ggctggtcat cctggacaga gtggtcagaa tgtaatgctc agtgtgggcg gggctggcag
     7441 agacggacac gcagctgcac caatccagca ccactcaatg gaggagcctt ctgtgatgga
     7501 ccgcccttcc agagagtcac ctgtaccacc ctctgtccag tggatggagg ctggaccgag
     7561 tgggccaagt ggtctgcgtg tgggacggag tgcacacatt ggcgcagtcg ggaatgtcag
     7621 gctccaccgc cacgtaatgg aggacgacac tgcagcggca gcatgatgga gagcaagaac
     7681 tgcactgagg gattatgtgc acgcaataaa aaggtttctg ttgaacatac aagtcatcct
     7741 ctgggctctg ggactggtgg cgcgcc
//
