LOCUS       sequence-44175-e        6606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  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 6606)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.diff.2012.06.004"; journalName: "Differentiation"; date: 
            "2012-09"; volume: "84"; issue: "2"; pages: "203-13"
COMMENT     Sequence Label: pcDNA-Wnt3
FEATURES             Location/Qualifiers
     source          1..6606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(2046..2070)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2096..2137
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2177..2196)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2221..2269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2471..2899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2558..2577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2768..2789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2908..2928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2913..3242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3093..3228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3309..4103
                     /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(3363..3382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3973..3992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4279..4400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4316..4335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4370..4389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4449..4465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4449..4465)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4462..4484)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4473..4489
                     /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(join(4497..4503,4504..4521,4522..4527))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4542..4563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4680..4697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4851..5439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4931..4950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5610..6401,6402..6470))
                     /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     6233..6252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6471..6575)
                     /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 tcaccatgga
      961 gccccacctg ctcgggctgc tcctcggcct cctgctcggt ggcaccaggg tcctcgctgg
     1021 ctacccaatt tggtggtccc tggccctggg ccagcagtac acatctctgg gctcacagcc
     1081 cctgctctgc ggctccatcc caggcctggt ccccaagcaa ctgcgcttct gccgcaatta
     1141 catcgagatc atgcccagcg tggccgaggg cgtgaagctg ggcatccagg agtgccagca
     1201 ccagttccgg ggccgccgct ggaactgcac caccatagat gacagcctgg ccatctttgg
     1261 gcccgtcctc gacaaagcca cccgcgagtc ggccttcgtt cacgccatcg cctcggccgg
     1321 cgtggccttc gccgtcaccc gctcctgcgc cgagggcacc tccaccattt gcggctgtga
     1381 ctcgcatcat aaggggccgc ctggcgaagg ctggaagtgg ggcggctgca gcgaggacgc
     1441 tgacttcggc gtgttagtgt ccagggagtt cgcggatgcg cgcgagaaca ggccggacgc
     1501 gcgctcggcc atgaacaagc acaacaacga ggcgggccgc acgactatcc tggaccacat
     1561 gcacctcaaa tgcaagtgcc acgggctgtc gggcagctgt gaggtgaaga cctgctggtg
     1621 ggcgcagcct gacttccgtg ccatcggtga cttcctcaag gacaagtatg acagcgcctc
     1681 ggagatggta gtagagaagc accgtgagtc ccgaggctgg gtggagaccc tccgggccaa
     1741 gtactcgctc ttcaagccac ccacggagag ggacctggtc tactacgaga actcccccaa
     1801 cttttgtgag cccaacccag agacgggttc ctttggcaca agggaccgga cttgcaatgt
     1861 cacctcccac ggcatcgatg gctgcgatct gctctgctgt ggccggggcc acaacacgag
     1921 gacggagaag cggaaggaaa aatgccactg catcttccac tggtgctgct acgtcagctg
     1981 ccaggagtgt attcgcatct acgacgtgca cacctgcaag tgaattctga agggtgggcg
     2041 cgccgaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa
     2101 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggttagt aatgagttta
     2161 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     2221 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     2281 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     2341 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     2401 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     2461 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     2641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2821 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     2881 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3061 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3121 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3181 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3241 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3481 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4081 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     4141 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4201 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4261 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4321 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4381 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4441 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4501 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4561 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4621 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4681 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4741 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4801 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5041 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5281 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5521 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5581 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5641 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5701 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5761 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5821 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5881 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5941 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6001 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6061 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6121 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6181 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6241 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6301 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6361 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6421 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6481 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6601 gacgtc
//
