LOCUS       sequence-373038-        6651 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 6651)
  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 6651)
  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-Wnt1
FEATURES             Location/Qualifiers
     source          1..6651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        280..659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        660..863
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     814..834
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     908..927
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        908..926
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    956..980
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2136..2160)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2186..2227
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2267..2286)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2311..2359
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2561..2989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2648..2667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2858..2879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2998..3018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3003..3332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3183..3318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3245..3264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3399..4193
                     /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(3453..3472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4063..4082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4369..4490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4406..4425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4460..4479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4539..4555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4539..4555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4552..4574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4563..4579
                     /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(4587..4593,4594..4611,4612..4617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4632..4653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4770..4787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4941..5529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5021..5040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5700..6491,6492..6560))
                     /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     6323..6342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6561..6651,1..14))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
       61 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      121 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
      181 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      241 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      301 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      361 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      421 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      481 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      541 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      601 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      661 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      721 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      781 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      841 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      901 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagttaagc tatcaacaag
      961 tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc atggggctct gggcgctgtt
     1021 gcctggctgg gtttctgcta cgctgctgct ggcgctggcc gctctgcccg cagccctggc
     1081 tgccaacagc agtggccgat ggtggggtat tgtgaacgta gcctcctcca cgaacctgct
     1141 tacagactcc aagagtctgc aactggtact cgagcccagt ctgcagctgt tgagccgcaa
     1201 acagcggcgt ctgatacgcc aaaatccggg gatcctgcac agcgtgagtg gggggctgca
     1261 gagtgccgtg cgcgagtgca agtggcagtt ccggaatcgc cgctggaact gtcccactgc
     1321 tccagggccc cacctcttcg gcaagatcgt caaccgaggc tgtcgagaaa cggcgtttat
     1381 cttcgctatc acctccgccg gggtcaccca ttcggtggcg cgctcctgct cagaaggttc
     1441 catcgaatcc tgcacgtgtg actaccggcg gcgcggcccc gggggccccg actggcactg
     1501 ggggggctgc agcgacaaca ttgacttcgg ccgcctcttc ggccgggagt tcgtggactc
     1561 cggggagaag gggcgggacc tgcgcttcct catgaacctt cacaacaacg aggcaggccg
     1621 tacgaccgta ttctccgaga tgcgccagga gtgcaagtgc cacgggatgt ccggctcatg
     1681 cacggtgcgc acgtgctgga tgcggctgcc cacgctgcgc gccgtgggcg atgtgctgcg
     1741 cgaccgcttc gacggcgcct cgcgcgtcct gtacggcaac cgcggcagca accgcgcttc
     1801 gcgggcggag ctgctgcgcc tggagccgga agacccggcc cacaaaccgc cctcccccca
     1861 cgacctcgtc tacttcgaga aatcgcccaa cttctgcacg tacagcggac gcctgggcac
     1921 agcaggcacg gcagggcgcg cctgtaacag ctcgtcgccc gcgctggacg gctgcgagct
     1981 gctctgctgc ggcaggggcc accgcacgcg cacgcagcgc gtcaccgagc gctgcaactg
     2041 caccttccac tggtgctgcc acgtcagctg ccgcaactgc acgcacacgc gcgtactgca
     2101 cgagtgtctg tgaattctga agggtgggcg cgccgaccca gctttcttgt acaaagtggt
     2161 tgatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     2221 ttctacgcgt accggttagt aatgagttta aacgggggag gctaactgaa acacggaagg
     2281 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     2341 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     2401 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     2461 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     2521 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2761 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2821 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2881 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2941 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3241 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3301 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3361 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3421 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3481 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3541 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3601 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3661 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3721 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3781 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3841 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3901 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3961 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4021 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4081 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4141 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4201 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4261 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4321 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4381 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4441 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4501 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4561 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4621 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4681 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4741 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4801 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4861 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4921 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4981 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5041 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5101 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5161 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5221 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5281 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5341 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5401 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5461 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5521 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5581 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5641 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5701 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5761 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5821 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5881 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5941 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6001 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6061 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6121 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6181 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6241 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6301 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6361 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6421 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6481 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6541 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6601 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca a
//
