LOCUS       sequence-131380-        8694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Unc5a with a C-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8694)
  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 8694)
  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 Unc5A-FLAG
FEATURES             Location/Qualifiers
     source          1..8694
                     /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             2715..2738
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2789..2844
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2843..2862)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5080..5098)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5166..5270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5271..5339,5340..6131)
                     /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(5489..5508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6302..6890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6791..6810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7044..7061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7178..7199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7214..7219,7220..7237,7238..7244)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7252..7268
                     /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     7257..7279
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7276..7292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7276..7292
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7563..7942
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7943..8146
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8097..8117
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8143..8167
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     8648..8667
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gcggccgcgc caccatggcc gtccggcccg gcctgtggcc agcgctcctg ggcatagtcc
       61 tcactgcctg gcttcgtggt tcgggtgccc agcagagtgc cacagtggcc aacccagtgc
      121 ctggtgccaa cccggacctg ctgccccact tcctggtaga gccggaggac gtgtacattg
      181 tcaagaacaa gcccgtgctg ctggtgtgca aggctgtgcc cgccacccag atcttcttca
      241 agtgcaacgg ggaatgggtt cgccaggtcg atcacgtcat tgaacgcagc actgacggca
      301 gcagcggatt gccaaccatg gaggtccgga tcaacgtatc aaggcagcag gtcgagaaag
      361 tgtttgggct ggaggagtac tggtgccagt gtgtggcatg gagctcctca ggaaccacca
      421 aaagccagaa ggcctacatc cggattgcct atttgcgcaa gaactttgag caggagccgc
      481 tggccaagga agtgtcactg gagcaaggca ttgtgctacc ttgtcgcccc ccggaaggaa
      541 tccccccagc tgaggtggag tggctccgaa atgaggacct cgtggacccc tccctcgacc
      601 ccaatgtgta catcacacgg gagcacagcc tagtcgtgcg gcaggcccgc ctggccgaca
      661 ctgccaacta cacctgcgtg gccaagaaca tcgtggcccg tcgccgaagc gcctctgcgg
      721 ccgtcattgt ttatgtgaac ggtgggtggt cgacgtggac cgagtggtcc gtctgcagtg
      781 ccagctgtgg gcgtggctgg cagaaacgga gccggagctg caccaacccg gcacctctca
      841 acgggggcgc cttctgtgag gggcagaatg tccagaaaac agcctgcgcc actctgtgcc
      901 cagtggatgg gagctggagc ccatggagta agtggtcagc ctgcgggctt gactgcaccc
      961 actggcggag ccgggagtgc tccgacccag cgccccgcaa cggaggtgag gagtgccggg
     1021 gtgctgacct ggacacccgc aactgtacca gtgacctctg cctgcacacc tcttccggcc
     1081 ccgaggacgt ggctctctac atcggcctcg tcgccgtggc cgtgtgcctc atcttgctgc
     1141 tgctggtcct cgtcctcatc tactgccgca agaaggaagg actggactca gacgtggctg
     1201 actcatccat ccttacctca ggcttccagc ctgtcagcat caagcccagc aaagcagaca
     1261 atccccatct gctcaccatc caaccggacc tcagcaccac cacgaccacc taccagggca
     1321 gcctgtgtcc ccggcaggat ggacccagcc ccaagttcca gctctctaat ggtcacctgc
     1381 tcagcccact gggcagtggc cgccatacgc tgcaccacag ctcccccacc tctgaggctg
     1441 aggacttcgt ctcccgcctc tccacccaaa actactttcg ttctctgccc cgcggtacca
     1501 gcaacatggc ctatgggacc ttcaacttcc tcgggggccg gctgatgatc cctaacacag
     1561 gaatcagcct cctcataccc ccggacgcca tcccccgagg aaagatctac gagatctacc
     1621 tcactctgca caagccagaa gacgtgaggt tgcccctagc tggctgtcag accctgctga
     1681 gtcctatcgt tagctgtggg cccccaggag tcctgctcac ccggccagtc atccttgcca
     1741 tggaccactg cggggagccc agtcccgaca gctggagcct gcgcctcaaa aagcagtcct
     1801 gtgagggcag ctgggaggac gtgctgcacc ttggtgagga gtcgccctct catctctact
     1861 actgccagct ggaggccggg gcctgctatg tcttcaccga gcagctaggc cgctttgccc
     1921 tggtgggaga ggccctcagc gtggctgcca ccaagcgcct caggctcctt ctgtttgccc
     1981 ctgtggcctg tacgtccctc gagtacaaca tccgagtgta ctgcctgcac gacacccacg
     2041 atgctctcaa ggaggtggtg cagctggaga agcagctggg tggacagctg atccaggagc
     2101 cccgtgtcct gcacttcaaa gacagttacc acaacctacg tctgtccatc cacgacgtgc
     2161 ccagctccct gtggaagagc aagctccttg tcagctacca ggagatccct ttttaccaca
     2221 tctggaatgg cactcagcag tatctgcact gcaccttcac cctggagcgc gtcaatgcca
     2281 gcaccagcga cctggcctgc aaggtgtggg tgtggcaggt ggagggagat ggacagagct
     2341 tcaacatcaa ctttaacatc actaaggaca cgaggtttgc tgaaatgctg gctctggaga
     2401 gtgaaggggg ggtcccagcc ctggtgggcc ccagtgcctt caagatcccc ttcctcattc
     2461 ggcaaaagat cattaccagc ctggacccac cctgcagccg gggcgccgac tggcgaactc
     2521 tagcccagaa acttcacctg gacagccatc ttagcttctt tgcctccaag cccagcccta
     2581 cagccatgat cctcaaccta tgggaggcgc ggcacttccc caacggcaac ctcggccagc
     2641 tggccgcagc tgtggccgga ctgggccagc cagatgctgg cctcttcacc gtgtcagagg
     2701 ccgagtgcgg cggcgattac aaggatgatg atgataagta atctagacac gtgattaatt
     2761 aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg
     2821 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc
     2881 gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg
     2941 actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg
     3001 tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg
     3061 ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt
     3121 ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg
     3181 tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc
     3241 aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa
     3301 gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac
     3361 gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag
     3421 catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag
     3481 gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt
     3541 ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc
     3601 agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata
     3661 actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga
     3721 cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat
     3781 ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata
     3841 tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca
     3901 cacgaattta tggctatggg caacacataa tcctagtgca atatgatact ggggttatta
     3961 agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac
     4021 aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc
     4081 gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt
     4141 ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg
     4201 gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca
     4261 aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg
     4321 cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc
     4381 tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg
     4441 gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4501 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4561 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     4621 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4681 atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata tgctatccta
     4741 atttatatct gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
     4801 atatctgggt agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     4861 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     4921 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     4981 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga
     5041 taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt
     5101 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5161 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5221 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5281 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5341 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5401 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5461 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     5521 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5581 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5641 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5701 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5761 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     5821 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5881 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5941 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6001 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6061 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6121 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6181 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6241 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6421 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6661 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6721 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6781 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6841 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6901 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6961 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7021 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7081 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7141 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7201 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7261 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg
     7321 tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt
     7381 gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt
     7441 agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac
     7501 gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg
     7561 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     7621 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7681 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7741 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7801 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     7861 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     7921 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     7981 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     8041 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     8101 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     8161 tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg gctcgcggtt
     8221 gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt cggcctccga
     8281 acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg gaaaacctct
     8341 cgagaaaggc gtctaaccag tcacagtcgc aaggtaggct gagcaccgtg gcgggcggca
     8401 gcgggtggcg gtcggggttg tttctggcgg aggtgctgct gatgatgtaa ttaaagtagg
     8461 cggtcttgag acggcggatg gtcgaggtga ggtgtggcag gcttgagatc cagctgttgg
     8521 ggtgagtact ccctctcaaa agcgggcatt acttctgcgc taagattgtc agtttccaaa
     8581 aacgaggagg atttgatatt cacctggccc gatctggcca tacacttgag tgacaatgac
     8641 atccactttg cctttctctc cacaggtgtc cactcccagg tccaagttta aact
//
