LOCUS       Exported                6603 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA-Wnt11-V5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6603)
  AUTHORS   Najdi R, Proffitt K, Sprowl S, Kaur S, Yu J, Covey TM, Virshup DM, 
            Waterman ML
  TITLE     A uniform human Wnt expression library reveals a shared secretory 
            pathway and unique signaling activities.
  JOURNAL   Differentiation. 2012 Sep;84(2):203-13. doi: 
            10.1016/j.diff.2012.06.004. Epub 2012 Jul 9.
  PUBMED    22784633
REFERENCE   2  (bases 1 to 6603)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6603
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2043..2067)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2093..2134
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2174..2193)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2218..2266
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2468..2896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2555..2574)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2765..2786
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2905..2925)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2910..3239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3090..3225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3152..3171
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3306..4100
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3360..3379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3970..3989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4276..4397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4313..4332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4367..4386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4446..4462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4446..4462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4459..4481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4470..4486
                     /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)."
     promoter        complement(4494..4524)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4539..4560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4677..4694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4848..5436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4928..4947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5607..6467)
                     /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     6230..6249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6468..6572)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgag
      961 ggcgcggccg caggtctgcg aggcgctgct cttcgccctg gcgctccaga ccggcgtgtg
     1021 ctatggcatc aagtggctgg cgctgtccaa gacaccatcg gccctggcac tgaaccagac
     1081 gcaacactgc aagcagctgg agggtctggt gtctgcacag gtgcagctgt gccgcagcaa
     1141 cctggagctc atgcacacgg tggtgcacgc cgcccgcgag gtcatgaagg cctgtcgccg
     1201 ggcctttgcc gacatgcgct ggaactgctc ctccattgag ctcgccccca actatttgct
     1261 tgacctggag agagggaccc gggagtcggc cttcgtgtat gcgctgtcgg ccgccgccat
     1321 cagccacgcc atcgcccggg cctgcacctc cggcgacctg cccggctgct cctgcggccc
     1381 cgtcccaggt gagccacccg ggcccgggaa ccgctgggga ggatgtgcgg acaacctcag
     1441 ctacgggctc ctcatggggg ccaagttttc cgatgctcct atgaaggtga aaaaaacagg
     1501 atcccaagcc aataaactga tgcgtctaca caacagtgaa gtggggagac aggctctgcg
     1561 cgcctctctg gaaatgaagt gtaagtgcca tggggtgtct ggctcctgct ccatccgcac
     1621 ctgctggaag gggctgcagg agctgcagga tgtggctgct gacctcaaga cccgatacct
     1681 gtcggccacc aaggtagtgc accgacccat gggcacccgc aagcacctgg tgcccaagga
     1741 cctggatatc cggcctgtga aggactcgga actcgtctat ctgcagagct cacctgactt
     1801 ctgcatgaag aatgagaagg tgggctccca cgggacacaa gacaggcagt gcaacaagac
     1861 atccaacgga agcgacagct gcgaccttat gtgctgcggg cgtggctaca acccctacac
     1921 agaccgcgtg gtcgagcggt gccactgtaa gtaccactgg tgctgctacg tcacctgccg
     1981 caggtgtgag cgtaccgtgg agcgctatgt ctgcaagtgc attctgaagg gtgggcgcgc
     2041 cgacccagct ttcttgtaca aagtggttga tctagagggc ccgcggttcg aaggtaagcc
     2101 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggttagtaat gagtttaaac
     2161 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     2221 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     2281 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     2341 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     2401 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     2461 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2521 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2581 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     2641 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2701 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2761 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2821 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2881 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2941 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3061 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3121 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3181 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3241 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3301 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3361 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3421 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3481 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3541 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3601 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3661 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3721 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3781 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3841 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3901 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3961 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4021 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4081 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     4141 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4201 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4261 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4321 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4381 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4441 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4501 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4681 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4741 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5041 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5281 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5821 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5881 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5941 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6001 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6061 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6121 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6181 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6241 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6301 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6361 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6421 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6481 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6541 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6601 gtc
//
