LOCUS       Exported                6594 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA-Wnt8A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  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 6594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6594
                     /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"
     CDS             1513..1524
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(2034..2058)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2084..2125
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2165..2184)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2209..2257
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2459..2887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2546..2565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2756..2777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2896..2916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2901..3230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3081..3216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3143..3162
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3297..4091
                     /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(3351..3370)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3961..3980
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4267..4388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4304..4323)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4358..4377
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4437..4453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4437..4453)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4450..4472)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4461..4477
                     /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(4485..4515)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4530..4551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4668..4685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4839..5427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4919..4938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5598..6458)
                     /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     6221..6240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6459..6563)
                     /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 tcaccatggg
      961 gaacctgttt atgctctggg cagctctggg catatgctgt gctgcattca gtgcctctgc
     1021 ctggtcagtg aacaatttcc tgataacagg tcccaaggcc tatctgacct acacgactag
     1081 tgtggccttg ggtgcccaga gtggcatcga ggagtgcaag ttccagtttg cttgggaacg
     1141 ctggaactgc cctgaaaatg ctcttcagct ctccacccac aacaggctga gaagtgctac
     1201 cagagagact tccttcatac atgctatcag ctctgctgga gtcatgtaca tcatcaccaa
     1261 gaactgtagc atgggtgact tcgaaaactg tggctgtgat gggtcaaaca atggaaaaac
     1321 aggaggccat ggctggatct ggggaggctg cagcgacaat gtggaatttg gggaaaggat
     1381 ctccaaactc tttgtggaca gtttggagaa ggggaaggat gccagagccc tgatgaatct
     1441 tcacaacaac agggccggca gactggcagt gagagccacc atgaaaagga catgcaaatg
     1501 tcatggcatc tctgggagct gcagcataca gacatgctgg ctgcagctgg ctgagttccg
     1561 ggagatggga gactacctaa aggccaagta tgaccaggcg ctgaaaattg aaatggataa
     1621 gcggcagctg agagctggga acagcgccga gggccactgg gtgcccgctg aggccttcct
     1681 tcctagcgca gaggcggaac tgatcttttt agaggaatca ccagattact gtacctgcaa
     1741 ttccagcctg ggcatctatg gcacagaggg tcgtgagtgc ctacagaaca gccacaacac
     1801 atccaggtgg gagcgacgta gctgtgggcg cctgtgcact gagtgtgggc tgcaggtgga
     1861 agagaggaaa actgaggtca taagcagctg taactgcaaa ttccagtggt gctgtacggt
     1921 caagtgtgac cagtgtaggc atgtggtgag caagtattac tgcgcacgct ccccaggcag
     1981 tgcccagtcc ctgggtaagg gcagtgcctg aattctgaag ggtgggcgcg ccgacccagc
     2041 tttcttgtac aaagtggttg atctagaggg cccgcggttc gaaggtaagc ctatccctaa
     2101 ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttaaa cgggggaggc
     2161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2221 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2281 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2341 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2401 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2581 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2821 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2881 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3241 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3721 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3841 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4081 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4261 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4381 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
