LOCUS       sequence-44186-e        6792 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 6792)
  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 6792)
  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-Wnt10A
FEATURES             Location/Qualifiers
     source          1..6792
                     /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(2232..2256)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2282..2323
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2363..2382)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2407..2455
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2657..3085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2744..2763)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2954..2975
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3094..3114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3099..3428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3279..3414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3341..3360
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3495..4289
                     /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(3549..3568)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4159..4178
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4465..4586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4502..4521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4556..4575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4635..4651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4635..4651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4648..4670)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4659..4675
                     /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(4683..4689,4690..4707,4708..4713))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4728..4749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4866..4883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5037..5625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5117..5136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5796..6587,6588..6656))
                     /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     6419..6438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6657..6761)
                     /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 cagcgcccac cctcgcccct ggctgcggct ccgaccccag ccccagccgc ggccagcgct
     1021 ctgggtgctc ctgttcttcc tactgctgct ggctgctgcc atgcccaggt cagcacccaa
     1081 tgacattctg gacctccgcc tccccccgga gcccgtgctc aatgccaaca cagtgtgcct
     1141 aacattgcca ggcctgagcc ggcggcagat ggaggtgtgt gtgcgtcacc ctgatgtggc
     1201 tgcctcagcc atacagggca tccagatcgc catccacgaa tgccaacacc aattcaggga
     1261 ccagcgctgg aactgctcaa gcctggagac tcgcaacaag atcccctatg agagtcccat
     1321 cttcagcaga ggtttccgag agagcgcttt tgcctacgcc atcgcagcag ctggcgtggt
     1381 gcacgccgtg tccaatgcgt gtgccctggg caaactgaag gcctgtggct gtgatgcgtc
     1441 ccggcgaggg gacgaggagg ccttccgtag gaagctgcac cgcttacaac tggatgcact
     1501 gcagcgtggt aagggcctga gccatggggt cccggaacac ccagccctgc ccacagccag
     1561 cccaggcctg caggactcct gggagtgggg cggctgcagc cccgacatgg gcttcgggga
     1621 gcgcttttct aaggactttc tggactcccg ggagcctcac agagacatcc acgcgagaat
     1681 gaggcttcac aacaaccgag ttgggaggca ggcagtgatg gagaacatgc ggcggaagtg
     1741 caagtgccac ggcacgtcag gcagctgcca gctcaagacg tgctggcagg tgacgcccga
     1801 gttccgcacc gtgggggcgc tgctgcgcag ccgcttccac cgcgccacgc tcatccggcc
     1861 gcacaaccgc aacggcggcc agctggagcc gggcccagcg ggggcaccct cgccggctcc
     1921 gggcgctccc gggccgcgcc gacgggccag ccccgccgac ctggtctact tcgaaaagtc
     1981 tcccgacttc tgcgagcgcg agccgcgcct ggactcggcg ggcaccgtgg gccgcctgtg
     2041 caacaagagc agcgccggct cggatggctg cggcagcatg tgctgcggcc gcggccacaa
     2101 catcctgcgc cagacgcgca gcgagcgctg ccactgccgc ttccactggt gctgtttcgt
     2161 ggtctgcgaa gagtgccgca tcaccgagtg ggtcagcgtc tgcaagtgaa ttctgaaggg
     2221 tgggcgcgcc gacccagctt tcttgtacaa agtggttgat ctagagggcc cgcggttcga
     2281 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gttagtaatg
     2341 agtttaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     2401 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     2461 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     2521 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     2581 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     2641 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     2821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2881 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2941 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3001 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3061 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3361 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3421 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3481 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3541 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3601 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3661 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3721 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3781 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3841 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3901 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3961 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4021 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4081 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4141 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4201 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4261 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     4321 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4381 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4441 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4501 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4561 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4621 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4681 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4741 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4801 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4861 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4921 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4981 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5041 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5101 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5161 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5221 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5281 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5341 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5401 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5461 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5521 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5581 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5641 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5701 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5761 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5821 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5881 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5941 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6001 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6061 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6121 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6181 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6241 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6301 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6361 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6421 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6481 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6541 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6601 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6661 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6721 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6781 ccacctgacg tc
//
