LOCUS       sequence-131245-        7490 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the Unc5D, isoform z 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 7490)
  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 7490)
  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: Unc5d.z-AP-His
FEATURES             Location/Qualifiers
     source          1..7490
                     /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             4070..4087
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4113..4234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4150..4169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4204..4223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4272..4289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4443..5031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4523..4542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5202..5993,5994..6062))
                     /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     5825..5844
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6063..6167)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6235..6253
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6291..6313)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6315..6335)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6320..6650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6501..6636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6563..6582
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6787..6806
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6873..7328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7010..7031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7222..7241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7454..7476
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7468..7485
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7469..7485
                     /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 tggggacagg
     1621 ggctgcagac ggcagccgcg gggcgcgccg gtggctccca tggctaggac tctttttctg
     1681 ggcggcgggg gctgcggctg cccgaggagc cgatggcagt gaaattcttc ccgactctat
     1741 cccatctgcc cctggaacgc tgcctcattt catcgaggag cccgaagatg catacatcat
     1801 caagagcaac ccgattgcac tcaggtgcaa agcaaggcca gccatgcaga tcttcttcaa
     1861 atgcaatggg gagtgggtcc atcagaatga acatgtgtcc gaggagagcc tggatgagag
     1921 ctcaggcttg aaggtccggg aagtgtttat caatgtcacc aggcagcaag tggaggactt
     1981 ccatgggcca gaggactact ggtgccagtg tgtagcgtgg agccacctgg gaacgtccaa
     2041 gagtaggaag gcatctgtgc gcatagccta tttacggaaa aactttgaac aagatccaca
     2101 aggcagggag gttcctattg aaggcatgat cgtgctgcat tgtcgcccgc cagagggagt
     2161 tccagcagca gaggtggaat ggctgaaaaa tgaggagccc atcgactccg aacaggatga
     2221 gaacattgac accagggctg accataactt gatcatcaga caggcccggc tgtcagactc
     2281 agggaattac acctgtatgg cagccaacat cgtggccaag aggaggagcc tctcagcaac
     2341 tgtggtggtc tacgtggatg gcagctggga agtgtggagt gaatggtcag tctgcagccc
     2401 agagtgtgaa catcttcgta tccgtgagtg cacagctcca cccccaagaa atgggggcaa
     2461 gttctgtgaa ggtctcagcc aggaatctga aaactgcaca gatggtctct gcattctaga
     2521 taaaaaacct cttcatgaaa taaaacccca aagatggagt aggagaggca ttgagaatgc
     2581 cagcactagt ggatcttccg gaatcatccc agttgaggag gagaacccgg acttctggaa
     2641 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     2701 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     2761 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     2821 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     2881 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     2941 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     3001 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     3061 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     3121 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     3181 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     3241 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3301 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     3361 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     3421 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     3481 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     3541 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     3601 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     3661 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     3721 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     3781 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     3841 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     3901 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     3961 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     4021 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtc atcatcacca
     4081 tcaccattaa tctagagatc ttattaaagc agaacttgtt tattgcagct tataatggtt
     4141 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4201 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact ctagctagac
     4261 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4321 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4381 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4441 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4501 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4561 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4621 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4681 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4741 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4801 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4861 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4921 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4981 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5041 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5101 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5161 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5221 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5281 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5341 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5401 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5461 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5521 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5581 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5641 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5701 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5761 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5821 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5881 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5941 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6001 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6061 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6121 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6181 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6241 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6301 atgcagctcc cggaggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     6361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6481 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6541 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     6601 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     6661 gagcttgtat atccattttc gtcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6721 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6781 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6841 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     6901 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6961 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     7021 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     7081 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     7141 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     7201 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     7261 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     7321 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     7381 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     7441 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
//
