LOCUS       sequence-131236-        7577 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the Unc5A, isoform a 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 7577)
  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 7577)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: Unc5a.a-AP-His
FEATURES             Location/Qualifiers
     source          1..7577
                     /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             4157..4174
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4200..4321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4237..4256)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4291..4310
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4359..4376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4530..5118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4610..4629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5289..6080,6081..6149))
                     /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     5912..5931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6150..6254)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6322..6340
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6378..6400)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6402..6422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6407..6737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6588..6723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6650..6669
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6874..6893
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6960..7415)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7097..7118)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7309..7328
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7541..7563
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7555..7572
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7556..7572
                     /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 tggccgtccg
     1621 gcccggcctg tggccagcgc tcctgggcat agtcctcact gcctggcttc gtggttcggg
     1681 tgcccagcag agtgccacag tggccaaccc agtgcctggt gccaacccgg acctgctgcc
     1741 ccacttcctg gtagagccgg aggacgtgta cattgtcaag aacaagcccg tgctgctggt
     1801 gtgcaaggct gtgcccgcca cccagatctt cttcaagtgc aacggggaat gggttcgcca
     1861 ggtcgatcac gtcattgaac gcagcactga cggcagcagc ggattgccaa ccatggaggt
     1921 ccggatcaac gtatcaaggc agcaggtcga gaaagtgttt gggctggagg agtactggtg
     1981 ccagtgtgtg gcatggagct cctcaggaac caccaaaagc cagaaggcct acatccggat
     2041 tgcctatttg cgcaagaact ttgagcagga gccgctggcc aaggaagtgt cactggagca
     2101 aggcattgtg ctaccttgtc gccccccgga aggaatcccc ccagctgagg tggagtggct
     2161 ccgaaatgag gacctcgtgg acccctccct cgaccccaat gtgtacatca cacgggagca
     2221 cagcctagtc gtgcggcagg cccgcctggc cgacactgcc aactacacct gcgtggccaa
     2281 gaacatcgtg gcccgtcgcc gaagcgcctc tgcggccgtc attgtttatg tgaacggtgg
     2341 gtggtcgacg tggaccgagt ggtccgtctg cagtgccagc tgtgggcgtg gctggcagaa
     2401 acggagccgg agctgcacca acccggcacc tctcaacggg ggcgccttct gtgaggggca
     2461 gaatgtccag aaaacagcct gcgccactct gtgcccagtg gatgggagct ggagcccatg
     2521 gagtaagtgg tcagcctgcg ggcttgactg cacccactgg cggagccggg agtgctccga
     2581 cccagcgccc cgcaacggag gtgaggagtg ccggggtgct gacctggaca cccgcaactg
     2641 taccagtgac ctctgcctgc acacctcttc cactagtgga tcttccggaa tcatcccagt
     2701 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     2761 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     2821 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     2881 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     2941 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     3001 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     3061 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     3121 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     3181 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     3241 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     3301 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     3361 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     3421 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     3481 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     3541 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     3601 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     3661 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     3721 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     3781 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     3841 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     3901 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     3961 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     4021 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     4081 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     4141 cgccgcgcac ccgggtcatc atcaccatca ccattaatct agagatctta ttaaagcaga
     4201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4321 atcatgtctg gtcgactcta gctagactct tccgcttcct cgctcactga ctcgctgcgc
     4381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4501 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4561 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4621 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4681 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4741 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4801 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4861 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4921 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4981 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5041 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5101 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5161 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5221 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5281 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5341 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5401 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5461 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5521 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5581 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5641 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5701 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5761 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5821 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5881 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5941 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6001 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6061 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6121 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6181 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6241 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6301 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6361 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg aggaatgtgt gtcagttagg
     6421 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     6481 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6541 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6601 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6661 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6721 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgtc ccggagacgg
     6781 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6841 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6901 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     6961 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     7021 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     7081 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7141 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     7201 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7261 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7321 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7381 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     7441 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7501 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     7561 acgacggcca gtgaatt
//
