LOCUS       sequence-211496-        8700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Unc5d with a C-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8700)
  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 8700)
  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: pTT3 Unc5D-FLAG
FEATURES             Location/Qualifiers
     source          1..8700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..18
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2721..2744
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2795..2850
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2849..2868)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5086..5104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5172..5276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5277..5345,5346..6137)
                     /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(5495..5514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6308..6896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6797..6816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7050..7067
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7184..7205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7220..7225,7226..7243,7244..7250)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7258..7274
                     /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     7263..7285
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7282..7298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7282..7298
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7569..7948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7949..8152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8103..8123
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8149..8173
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     8654..8673
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gcggccgcgc caccatgggg acaggggctg cagacggcag ccgcggggcg cgccggtggc
       61 tcccatggct aggactcttt ttctgggcgg cgggggctgc ggctgcccga ggagccgatg
      121 gcagtgaaat tcttcccgac tctatcccat ctgcccctgg aacgctgcct catttcatcg
      181 aggagcccga agatgcatac atcatcaaga gcaacccgat tgcactcagg tgcaaagcaa
      241 ggccagccat gcagatcttc ttcaaatgca atggggagtg ggtccatcag aatgaacatg
      301 tgtccgagga gagcctggat gagagctcag gcttgaaggt ccgggaagtg tttatcaatg
      361 tcaccaggca gcaagtggag gacttccatg ggccagagga ctactggtgc cagtgtgtag
      421 cgtggagcca cctgggaacg tccaagagta ggaaggcatc tgtgcgcata gcctatttac
      481 ggaaaaactt tgaacaagat ccacaaggca gggaggttcc tattgaaggc atgatcgtgc
      541 tgcattgtcg cccgccagag ggagttccag cagcagaggt ggaatggctg aaaaatgagg
      601 agcccatcga ctccgaacag gatgagaaca ttgacaccag ggctgaccat aacttgatca
      661 tcagacaggc ccggctgtca gactcaggga attacacctg tatggcagcc aacatcgtgg
      721 ccaagaggag gagcctctca gcaactgtgg tggtctacgt ggatggcagc tgggaagtgt
      781 ggagtgaatg gtcagtctgc agcccagagt gtgaacatct tcgtatccgt gagtgcacag
      841 ctccaccccc aagaaatggg ggcaagttct gtgaaggtct cagccaggaa tctgaaaact
      901 gcacagatgg tctctgcatt ctagataaaa aacctcttca tgaaataaaa ccccaaagat
      961 ggagtaggag aggcattgag aatgccagcg acattgcttt gtactccggc ttgggtgcag
     1021 ccgtggtagc tgttgcggtc ctggtcattg gtgtcaccct ctacagacgg agccacagtg
     1081 actacggagt ggatgtcatt gactcttctg cattgacagg tggcttccag accttcaact
     1141 tcaaaacagt tcgtcaagga aattctctac tactgaatcc tgccatgcag ccagatctga
     1201 ctgtgagccg gacatacagc gggcccatct gtctgcagga tccgctcgac aaggagctca
     1261 tgacagagtc ctcactcttc aatcctttat ctgacatcaa agtcaaagtc cagagctcat
     1321 ttatggtttc cctgggagtg tctgagagag cggagtacca tggcaagaat cactcgggga
     1381 ctttccccca tggaaacaac cgcggattca gtacaataca tcccagaaac aaaacgccgt
     1441 acatccaaaa tctgtcatca ctgccgacaa ggacagagct gaggacaact ggtgtctttg
     1501 gccatttagg gggacgctta gtaatgccaa atacaggggt gagtctactc ataccacatg
     1561 gtgccatccc agaggagaat tcttgggaga tttatatgtc aatcaaccaa ggtgaaccga
     1621 gcctgcagtc agatggatct gaggttctcc tgagtcctga agtcacctgt gggcctccag
     1681 atatgcttgt cacaactccc tttgcgctga ccatccctca ctgtgcagac gtcagttcag
     1741 agcactggaa cattcacctg aagaagagga cccagcaggg gaagtgggag gaagtgatgt
     1801 cagtggagga tgagtccaca tcctgttact gcctcttgga tccctttgcc tgtcacgtgc
     1861 tgcttgatag ctttggaacg tatgccctca caggagagcc aatcacagac tgtgccgtaa
     1921 agcagctcaa ggtggctgtg tttggctgca tgtcctgtaa ctccttggat tacaacctga
     1981 gagtttactg tgtggacaat actccttgtg catttcagga agtgatttct gacgaacggc
     2041 accaaggcgg ccagctcctg gaagaaccaa agctgctgca tttcaaaggg aatacattca
     2101 gtctccaggt ctctgtcctt gacatccctc ccttcctctg gaggatcaaa ccgttcactg
     2161 catgccagga agtccccttc tcccgagtgt ggagcagcaa ccggcagccc ctgcactgtg
     2221 ccttctccct ggagcgctac acgcccacca ccacccagct gtcctgcaaa atctgcattc
     2281 ggcagctcaa aggccatgaa cagatcctcc aagtgcagac atccatccta gagagtgaac
     2341 gagaaaccat cactttcttc gcacaagagg acagtacctt tcctgcacag actggcccaa
     2401 aagccttcaa aattccctat tccatcagac agaggatctg cgccacgttt gataccccaa
     2461 atgccaaagg caaggactgg cagatgttag cacagaagaa cagcatcaat aggaatttat
     2521 catatttcgc tacccaaagc agcccttccg ctgtcatttt gaacctctgg gaagctcgtc
     2581 atcagcagga tggggacctc gactcgctcg cctgtgccct tgaggagatt gggaggacac
     2641 acacgaaact atccaacatc acagagccac agattgatga tgctgacttc aactacagca
     2701 ggcaaaatgg actcggcggc gattacaagg atgatgatga taagtaaact agacacgtga
     2761 ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat tttcattgca
     2821 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     2881 ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc ggacgaacta
     2941 aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat cccttcagtt
     3001 ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg ggaccaaaca
     3061 caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca tttgccaaca
     3121 ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca caacattgcc
     3181 tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg tacctcccag
     3241 gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct gtgtagctac
     3301 cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc cttgttaacc
     3361 ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt
     3421 caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt
     3481 cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt acatggggtc
     3541 aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga agatcaagga
     3601 gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg tttagctaat
     3661 agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc
     3721 aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt gtgctatgac
     3781 accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa tgaaacattc
     3841 tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga ctggatgtcc
     3901 atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat gatactgggg
     3961 ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt acactctatt
     4021 tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt tgtctctaac
     4081 acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag acaagtggcc
     4141 actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg ccgccctgcg
     4201 gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg ctaaccactg
     4261 cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc tgcataagta
     4321 ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact
     4381 tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca
     4441 cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat agcatatgct
     4501 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4561 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     4621 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4681 atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct
     4741 atcctaattt atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc
     4801 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc
     4861 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     4921 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     4981 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     5041 tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc gtgatacgcc
     5101 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5161 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5221 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5281 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5341 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5401 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5461 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     5521 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5581 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5641 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5701 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5761 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5821 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5881 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5941 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6001 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6061 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6121 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6181 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6241 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6421 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6721 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6901 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6961 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7021 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     7141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     7201 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     7261 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc
     7321 tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg caacgttgtt
     7381 gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga atcaatattg
     7441 gcaattagcc atattagtca ttggttatat agcataaatc aatattggct attggccatt
     7501 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
     7561 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7621 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7681 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7741 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7801 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     7861 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     7921 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     7981 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     8041 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     8101 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     8161 gaaccgtcag atcctcactc tcttccgcat cgctgtctgc gagggccagc tgttgggctc
     8221 gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa acccgtcggc
     8281 ctccgaacgg tactccgcca ccgagggacc tgagcgagtc cgcatcgacc ggatcggaaa
     8341 acctctcgag aaaggcgtct aaccagtcac agtcgcaagg taggctgagc accgtggcgg
     8401 gcggcagcgg gtggcggtcg gggttgtttc tggcggaggt gctgctgatg atgtaattaa
     8461 agtaggcggt cttgagacgg cggatggtcg aggtgaggtg tggcaggctt gagatccagc
     8521 tgttggggtg agtactccct ctcaaaagcg ggcattactt ctgcgctaag attgtcagtt
     8581 tccaaaaacg aggaggattt gatattcacc tggcccgatc tggccataca cttgagtgac
     8641 aatgacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca agtttaaact
//
