LOCUS       sequence-44187-e        6708 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6708)
  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 6708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Wnt10B
FEATURES             Location/Qualifiers
     source          1..6708
                     /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(2148..2172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2198..2239
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2279..2298)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2323..2371
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2573..3001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2660..2679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2870..2891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3010..3030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3015..3344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3195..3330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3257..3276
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3411..4205
                     /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(3465..3484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4075..4094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4381..4502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4418..4437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4472..4491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4551..4567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4551..4567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4564..4586)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4575..4591
                     /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(4599..4605,4606..4623,4624..4629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4644..4665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4782..4799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4953..5541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5033..5052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5712..6503,6504..6572))
                     /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     6335..6354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6573..6677)
                     /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 tcaccatgct
      961 ggaggagccc cggccgcggc ctccgccctc gggcctcgcg ggtctcctgt tcctggcgtt
     1021 gtgcagtcgg gctctaagca atgagattct gggcctgaag ttgcctggcg agccgccgct
     1081 gacggccaac accgtgtgct tgacgctgtc cggcctgagc aagcggcagc taggcctgtg
     1141 cctgcgcaac cccgacgtga cggcgtccgc gcttcagggt ctgcacatcg cggtccacga
     1201 gtgtcagcac cagctgcgcg accagcgctg gaactgctcc gcgcttgagg gcggcggccg
     1261 cctgccgcac cacagcgcca tcctcaagcg cggtttccga gaaagtgctt tttccttctc
     1321 catgctggct gctggggtca tgcacgcagt agccacggcc tgcagcctgg gcaagctggt
     1381 gagctgtggc tgtggctgga agggcagtgg tgagcaggat cggctgaggg ccaaactgct
     1441 gcagctgcag gcactgtccc gaggcaagag tttcccccac tctctgccca gccctggccc
     1501 tggctcaagc cccagccctg gcccccagga cacatgggaa tggggtggct gtaaccatga
     1561 catggacttt ggagagaagt tctctcggga tttcttggat tccagggaag ctccccggga
     1621 catccaggca cgaatgcgaa tccacaacaa cagggtgggg cgccaggtgg taactgaaaa
     1681 cctgaagcgg aaatgcaagt gtcatggcac atcaggcagc tgccagttca agacatgctg
     1741 gagggcggcc ccagagttcc gggcagtggg ggcggcgttg agggagcggc tgggccgggc
     1801 catcttcatt gatacccaca accgcaattc tggagccttc cagccccgtc tgcgtccccg
     1861 tcgcctctca ggagagctgg tctactttga gaagtctcct gacttctgtg agcgagaccc
     1921 cactatgggc tccccaggga caaggggccg ggcctgcaac aagaccagcc gcctgttgga
     1981 tggctgtggc agcctgtgct gtggccgtgg gcacaacgtg ctccggcaga cacgagttga
     2041 gcgctgccat tgccgcttcc actggtgctg ctatgtgctg tgtgatgagt gcaaggttac
     2101 agagtgggtg aatgtgtgta agtgaattct gaagggtggg cgcgccgacc cagctttctt
     2161 gtacaaagtg gttgatctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     2221 cctcggtctc gattctacgc gtaccggtta gtaatgagtt taaacggggg aggctaactg
     2281 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     2341 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     2401 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     2461 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     2521 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     2581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2701 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     2761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2941 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3001 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3361 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4201 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     4261 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4321 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4501 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4561 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4621 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4681 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4741 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4801 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4861 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4921 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4981 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5161 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5281 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5341 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5401 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5461 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5521 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5581 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5641 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5701 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5761 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5821 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5881 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5941 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6001 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6061 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6121 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6181 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6241 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6301 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6361 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6421 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6481 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6541 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6601 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6661 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
