LOCUS       Exported                6618 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA-Wnt5B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6618)
  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 6618)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6618
                     /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(2058..2082)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2108..2149
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2189..2208)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2233..2281
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2483..2911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2570..2589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2780..2801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2920..2940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2925..3254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3105..3240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3167..3186
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3321..4115
                     /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(3375..3394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3985..4004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4291..4412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4328..4347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4382..4401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4461..4477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4461..4477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4474..4496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4485..4501
                     /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(4509..4539)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4554..4575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4692..4709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4863..5451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4943..4962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5622..6482)
                     /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     6245..6264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6483..6587)
                     /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 tcaccatgcc
      961 cagcctgctg ctgctgttca cggctgctct gctgtccagc tgggctcagc ttctgacaga
     1021 cgccaactcc tggtggtcat tagctttgaa cccggtgcag agacccgaga tgtttatcat
     1081 cggtgcccag cccgtgtgca gtcagcttcc cgggctctcc cctggccaga ggaagctgtg
     1141 ccaattgtac caggagcaca tggcctacat aggggaggga gccaagactg gcatcaagga
     1201 atgccagcac cagttccggc agcggcggtg gaattgcagc acagcggaca acgcatctgt
     1261 ctttgggaga gtcatgcaga taggcagccg agagaccgcc ttcacccacg cggtgagcgc
     1321 cgcgggcgtg gtcaacgcca tcagccgggc ctgccgcgag ggcgagctct ccacctgcgg
     1381 ctgcagccgg acggcgcggc ccaaggacct gccccgggac tggctgtggg gcggctgtgg
     1441 ggacaacgtg gagtacggct accgcttcgc caaggagttt gtggatgccc gggagcgaga
     1501 gaagaacttt gccaaaggat cagaggagca gggccgggtg ctcatgaacc tgcaaaacaa
     1561 cgaggccggt cgcagggctg tgtataagat ggcagacgta gcctgcaaat gccacggcgt
     1621 ctcggggtcc tgcagcctca agacctgctg gctgcagctg gccgagttcc gcaaggtcgg
     1681 ggaccggctg aaggagaagt acgacagcgc ggccgccatg cgcgtcaccc gcaagggccg
     1741 gctggagctg gtcaacagcc gcttcaccca gcccaccccg gaggacctgg tctatgtgga
     1801 ccccagcccc gactactgcc tgcgcaacga gagcacgggc tccctgggca cgcagggccg
     1861 cctctgcaac aagacctcgg agggcatgga tggctgtgag ctcatgtgct gcgggcgtgg
     1921 ctacaaccag ttcaagagcg tgcaggtgga gcgctgccac tgcaagttcc actggtgctg
     1981 cttcgtcagg tgtaagaagt gcacggagat cgtggaccag tacatctgta aatgaattct
     2041 gaagggtggg cgcgccgacc cagctttctt gtacaaagtg gttgatctag agggcccgcg
     2101 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtta
     2161 gtaatgagtt taaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2221 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     2281 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2341 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2401 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     2461 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     2641 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2821 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     2881 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3001 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3061 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3181 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3241 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3301 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3361 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3421 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3481 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3541 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3601 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3661 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3721 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3781 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3841 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3901 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3961 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4021 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4081 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4141 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4201 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4261 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4321 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4381 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4441 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4501 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4561 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4621 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4681 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5041 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5281 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5641 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5701 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5761 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5821 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5881 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5941 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6001 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6061 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6121 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6181 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6241 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6301 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6361 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6421 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6481 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6541 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6601 aaagtgccac ctgacgtc
//
