LOCUS       sequence-199981-        7959 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 7959)
  AUTHORS   Shimizu H, Julius MA, Giarre M, Zheng Z, Brown AM, Kitajewski J
  TITLE     Transformation by Wnt family proteins correlates with regulation of 
            beta-catenin.
  JOURNAL   Cell Growth Differ. 1997 Dec . 8(12):1349-58.
  PUBMED    9419423
REFERENCE   2  (bases 1 to 7959)
  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"; journalName: "Cell Growth
            Differ. 1997 Dec . 8(12):1349-58."
COMMENT     Sequence Label: pLNC Wnt-3aHA
FEATURES             Location/Qualifiers
     source          1..7959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     704..720
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     704..720
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1927..1953
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1933..1959
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     LTR             2087..2680
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2783..2805)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2891..3031
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2946..2965
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3046..3063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3217..3805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3297..3316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3976..4767,4768..4836))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4599..4618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4837..4941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5009..5027
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5599..5774
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5837..6036
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6237..6653
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6583..6605
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6621..6637
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6697..7491
                     /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(6751..6770)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7361..7380
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ttaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggcccc aagctgcggc cgctctagaa ctagtggatc
      721 ccccgggctg caggaattca tgtcttacgg tcaaggcaga gggcccagcg ccactgcagc
      781 cgcgccacct cccagggccg ggccagccca ggcgtccgcg ctctcggggt ggactccccc
      841 cgctgcgcgc tcaagccggc gatggctcct ctcggatacc tcttagtgct ctgcagcctg
      901 aagcaggctc tgggcagcta cccgatctgg tggtccttgg ctgtgggacc ccagtactcc
      961 tctctgagca ctcagcccat tctctgtgcc agcatcccag gcctggtacc gaagcagctg
     1021 cgcttctgca ggaactacgt ggagatcatg cccagcgtgg ctgagggtgt caaagcgggc
     1081 atccaggagt gccagcacca gttccgaggc cggcgttgga actgcaccac cgtcagcaac
     1141 agcctggcca tctttggccc tgttctggac aaagccaccc gggagtcagc ctttgtccat
     1201 gccatcgcct ccgctggagt agctttcgca gtgacacgct cctgtgcaga gggatcagct
     1261 gctatctgtg ggtgcagcag ccgcctccag ggctccccag gcgagggctg gaagtggggc
     1321 ggctgtagtg aggacattga atttggagga atggtctctc gggagtttgc cgatgccagg
     1381 gagaaccggc cggatgcccg ctctgccatg aaccgtcaca acaatgaggc tgggcgccag
     1441 gccatcgcca gtcacatgca cctcaagtgc aaatgccacg ggctatctgg cagctgtgaa
     1501 gtgaagacct gctggtggtc gcagccggac ttccgcacca tcggggattt cctcaaggac
     1561 aagtatgaca gtgcctcgga gatggtggta gagaaacacc gagagtctcg tggctgggtg
     1621 gagaccctga ggccacgtta cacgtacttc aaggtgccga cagaacgcga cctggtctac
     1681 tacgaggcct cacccaactt ctgcgaacct aaccccgaaa ccggctcctt cgggacgcgt
     1741 gaccgcacct gcaatgtgag ctcgcatggc atagatgggt gcgacctgtt gtgctgcggg
     1801 cgcgggcata acgcgcgcac tgagcgacgg agggagaaat gccactgtgt tttccattgg
     1861 tgctgctacg tcagctgcca ggagtgcaca cgtgtctatg acgtgcacac ctgcaagtcc
     1921 atggcctacc catatgatgt tccagattac gcttctttgg gtccagggcc ttaattaatt
     1981 aagatccggc tctagaggat ccgtcgacct cgaggggggg cccggtaccc taacacgagc
     2041 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     2101 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     2161 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     2221 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     2281 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     2341 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     2401 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     2461 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     2521 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     2581 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     2641 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     2701 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc
     2761 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2821 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2881 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     2941 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     3001 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     3061 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3121 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3181 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3241 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3301 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3361 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3421 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3481 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3541 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3601 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3661 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3721 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3781 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3961 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4021 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4081 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4141 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4201 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4261 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     4321 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4381 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4441 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4501 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4561 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     4621 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4681 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4741 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4801 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4861 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4921 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4981 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     5041 agaattcata ccagatcacc gaaaactgtc ctccaaatgt gtccccctca cactcccaaa
     5101 ttcgcgggct tctgcctctt agaccactct accctattcc ccacactcac cggagccaaa
     5161 gccgcggccc ttccgtttct ttgcttttga aagaccccac ccgtaggtgg caagctagct
     5221 taagtaacgc cactttgcaa ggcatggaaa aatacataac tgagaataga aaagttcaga
     5281 tcaaggtcag gaacaaagaa acagctgaat accaaacagg atatctgtgg taagcggttc
     5341 ctgccccggc tcagggccaa gaacagatga gacagctgag tgatgggcca aacaggatat
     5401 ctgtggtaag cagttcctgc cccggctcgg ggccaagaac agatggtccc cagatgcggt
     5461 ccagccctca gcagtttcta gtgaatcatc agatgtttcc agggtgcccc aaggacctga
     5521 aaatgaccct gtaccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     5581 gcttccgctc tccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt
     5641 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     5701 cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac
     5761 gggggtcttt catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga
     5821 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
     5881 ctatgtttga tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     5941 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     6001 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     6061 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     6121 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     6181 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     6241 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     6301 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     6361 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     6421 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     6481 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     6541 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     6601 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaattc
     6661 cgatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6721 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6781 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6841 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6901 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6961 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     7021 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     7081 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7141 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7201 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7261 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7321 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7381 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7441 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7501 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     7561 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     7621 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg
     7681 cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca
     7741 aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc
     7801 aggagtgggg aggcacgatg gccgctttgg tcgaggcgga tccggccatt agccatatta
     7861 ttcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcca
     7921 tatcataata tgtacattta tattggctca tgtccaaca
//
