LOCUS       sequence-44191-e        6621 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 6621)
  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 6621)
  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-Wnt2-V5
FEATURES             Location/Qualifiers
     source          1..6621
                     /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(2061..2085)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2111..2152
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2192..2211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2236..2284
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2486..2914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2783..2804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2923..2943)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2928..3257
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3108..3243
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3170..3189
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3324..4118
                     /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(3378..3397)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3988..4007
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4294..4415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4331..4350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4385..4404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4464..4480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4464..4480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4477..4499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4488..4504
                     /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(4512..4518,4519..4536,4537..4542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4557..4578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4695..4712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4866..5454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4946..4965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5625..6416,6417..6485))
                     /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     6248..6267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6486..6590)
                     /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 tcaccatgaa
      961 cgcccctctc ggtggaatct ggctctggct ccctctgctc ttgacctggc tcacccccga
     1021 ggtcaactct tcatggtggt acatgagagc tacaggtggc tcctccaggg tgatgtgcga
     1081 taatgtgcca ggcctggtga gcagccagcg gcagctgtgt caccgacatc cagatgtgat
     1141 gcgtgccatt agccagggcg tggccgagtg gacagcagaa tgccagcacc agttccgcca
     1201 gcaccgctgg aattgcaaca ccctggacag ggatcacagc ctttttggca gggtcctact
     1261 ccgaagtagt cgggaatctg cctttgttta tgccatctcc tcagctggag ttgtatttgc
     1321 catcaccagg gcctgtagcc aaggagaagt aaaatcctgt tcctgtgatc caaagaagat
     1381 gggaagcgcc aaggacagca aaggcatttt tgattggggt ggctgcagtg ataacattga
     1441 ctatgggatc aaatttgccc gcgcatttgt ggatgcaaag gaaaggaaag gaaaggatgc
     1501 cagagccctg atgaatcttc acaacaacag agctggcagg aaggctgtaa agcggttctt
     1561 gaaacaagag tgcaagtgcc acggggtgag cggctcatgt actctcagga catgctggct
     1621 ggccatggcc gacttcagga aaacgggcga ttatctctgg aggaagtaca atggggccat
     1681 ccaggtggtc atgaaccagg atggcacagg tttcactgtg gctaacgaga ggtttaagaa
     1741 gccaacgaaa aatgacctcg tgtattttga aaattctcca gactactgta tcagggaccg
     1801 agaggcaggc tccctgggta cagcaggccg tgtgtgcaac ctgacttccc ggggcatgga
     1861 cagctgtgaa gtcatgtgct gtgggagagg ctacgacacc tcccatgtca cccggatgac
     1921 caagtgtggg tgtaagttcc actggtgctg cgccgtgcgc tgtcaggact gcctggaagc
     1981 tctggatgtg cacacatgca aggcccccaa gaacgctgac tggacaaccg ctacatgcat
     2041 tctgaagggt gggcgcgccg acccagcttt cttgtacaaa gtggttgatc tagagggccc
     2101 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     2161 ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     2221 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     2281 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     2341 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     2401 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     2461 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2821 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     2881 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2941 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3001 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3121 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3181 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3241 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3301 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3361 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3421 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3481 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3541 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3601 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3661 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3721 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3781 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3841 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3901 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3961 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4021 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4081 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     4141 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4201 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4261 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4381 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4441 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4501 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4561 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4621 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4801 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5041 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5281 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5341 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5401 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5461 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5521 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5581 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5641 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5701 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5761 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5821 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5881 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5941 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6001 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6061 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6121 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6181 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6241 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6301 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6361 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6421 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6481 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6541 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6601 cgaaaagtgc cacctgacgt c
//
