LOCUS       sequence-131216-        8510 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the Sema4C 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 8510)
  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 8510)
  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: Sema4c-AP-His
FEATURES             Location/Qualifiers
     source          1..8510
                     /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             5090..5107
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5133..5254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5170..5189)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5224..5243
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5292..5309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5463..6051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5543..5562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6222..7013,7014..7082))
                     /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     6845..6864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7083..7187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7255..7273
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7311..7333)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7335..7355)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7340..7670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7521..7656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7583..7602
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7807..7826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7893..8348)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8030..8051)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8242..8261
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8474..8496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8488..8505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8489..8505
                     /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 tggccccaca
     1621 ctgggctgtc tggctgctgg cagcagggct gtggggcctg ggcatcgggg ctgagatgtg
     1681 gtggaacctt gtgccccgga agacagtatc ttctggggag ctggtcacag tagtgaggcg
     1741 gttctcccag acaggcatcc aggacttcct gacactgacc ctgacagaac attctggcct
     1801 tttatatgtg ggggcccgag aggcgctgtt tgccttcagt gtagaggctc tggagctgca
     1861 aggagcgatc tcttgggagg ctccagctga gaagaaaatt gaatgtaccc agaaagggaa
     1921 gagcaaccag accgaatgct tcaacttcat ccgcttcctt cagccataca attcctccca
     1981 tctgtatgtc tgcggcacct atgccttcca gcccaagtgc acctacatca acatgctcac
     2041 gttcaccttg gaccgtgcag aatttgagga tgggaagggt aaatgcccat atgacccagc
     2101 taagggtcac accggactcc ttgtggacgg tgagctgtac tcagccacac tcaataactt
     2161 cctgggcaca gagccggtta tccttcgata catggggacc caccactcca tcaagacaga
     2221 gtacctggct ttttggctga atgaacccca ctttgtaggc tctgcctttg tccctgagag
     2281 tgtgggaagc ttcacgggag acgatgacaa gatctacttc ttcttcagtg agcgggcagt
     2341 ggagtatgac tgctattccg agcaggtggt ggctcgtgtg gcgagagtct gtaagggtga
     2401 catgggggga gcacggacgc tgcagaagaa atggacgacg ttcctgaagg ctcggttggt
     2461 gtgctcagcc cctgactgga aggtctactt caaccagctg aaggcggtgc acaccctgcg
     2521 gggcgcctct tggcacaaca ccaccttctt cggggttttt caagcgcgat ggggcgatat
     2581 ggacctgtct gcagtttgtg agtaccagtt ggaacagatc cagcaagtgt ttgagggtcc
     2641 ctacaaggag tacagtgagc aagcccagaa gtgggcccgc tatactgacc cggtacccag
     2701 ccctcggcct ggttcgtgta tcaacaactg gcaccgagac aatggctaca ccagttccct
     2761 ggaactgccg gacaacaccc tcaacttcat caagaagcac cccctgatgg aggaccaggt
     2821 gaagcctcgg ttgggccgcc ccctacttgt gaagaagaac actaacttca cacacgtggt
     2881 ggccgacagg gtcccagggc ttgatggtgc cacctataca gtgttgttca ttggtacagg
     2941 agatggctgg ctgctgaagg ctgtgagcct ggggccctgg atccacatgg tggaggaact
     3001 gcaggtgttt gaccaggagc cagtggaaag tctggtgctg tctcagagca agaaggtgct
     3061 ctttgctggc tcccgctctc agctggttca gctgtctctg gccgactgca caaagtaccg
     3121 tttctgtgta gactgtgtcc tggccaggga cccttactgt gcctggaatg tcaacaccag
     3181 ccgctgtgtg gccaccacca gtggtcgctc ggggtccttt ctggtccaac atgtggcgaa
     3241 cttggacact tcaaagatgt gtaaccagta tggcattaaa aaagtcagat ctattcccaa
     3301 gaacatcacc gttgtgtcag gcacagacct ggtcctaccc tgccacctct cgtccaattt
     3361 ggcccatgcc cactggacct tcggaagcca ggacctgcct gcagaacaac ctggctcctt
     3421 tctttatgac acgggactcc aggcgctggt ggtgatggcc gcacagtccc gtcactctgg
     3481 accctatcgt tgctattcag aggagcaggg gacaagactg gctgcagaaa gctaccttgt
     3541 tgctgtcgtg gccggctcgt cggtgacact ggaggcacgg gctcccttgg aaaacctggg
     3601 gggcgcgcct ggatcttccg gaatcatccc agttgaggag gagaacccgg acttctggaa
     3661 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     3721 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     3781 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     3841 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     3901 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     3961 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     4021 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     4081 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     4141 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     4201 catggacatt gacgtgatcc taggtggggg ccgaaagtac atgtttcgca tgggaacccc
     4261 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     4321 ggtgcaggaa tggctggcga agcgccaggg tgcccggtac gtgtggaacc gcactgagct
     4381 catgcaggct tccctggacc cgtctgtggc ccatctcatg ggtctctttg agcctggaga
     4441 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     4501 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     4561 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     4621 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     4681 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     4741 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     4801 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     4861 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     4921 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     4981 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     5041 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtc atcatcacca
     5101 tcaccattaa tctagagatc ttattaaagc agaacttgtt tattgcagct tataatggtt
     5161 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5221 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact ctagctagac
     5281 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6301 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6361 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6421 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6481 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6541 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6601 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6661 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6721 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6781 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6841 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6901 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6961 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7021 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7081 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7141 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7201 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     7261 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     7321 atgcagctcc cggaggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     7381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     7441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7501 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     7561 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     7621 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     7681 gagcttgtat atccattttc gtcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     7741 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     7801 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     7861 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     7921 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     7981 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     8041 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     8101 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     8161 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     8221 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     8281 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     8341 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8401 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8461 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
//
