LOCUS       sequence-156322-        8705 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 8705)
  AUTHORS   Kolligs FT, Hu G, Dang CV, Fearon ER
  TITLE     Neoplastic transformation of RK3E by mutant beta-catenin requires 
            deregulation of Tcf/Lef transcription but not activation of c-myc 
            expression.
  JOURNAL   Mol Cell Biol. 1999 Aug . 19(8):5696-706.
  PUBMED    10409758
REFERENCE   2  (bases 1 to 8705)
  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: "Mol Cell 
            Biol. 1999 Aug . 19(8):5696-706."
COMMENT     Sequence Label: pPGS dnTcf-4(deltaN41)
FEATURES             Location/Qualifiers
     source          1..8705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        497..876
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        877..1080
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1031..1051
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1319..1676
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1740..2156
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2086..2108
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2124..2140
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2166..2540
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2478..2494
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     primer_bind     2550..2570
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2552..2570
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2617..2640
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4322..4888
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4338..4890
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4482..4499)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4709..4728
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4890..5684
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4944..4963)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5554..5573
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             5874..6343
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6473..6490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6644..7232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6724..6743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7709..8500,8501..8569))
                     /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     8332..8351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8570..8674)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcttaggag tttcctaata catcccaaac tcaaatatat aaagcatttg acttgttcta
       61 tgccctaggg ggcgggggga agctaagcca gcttttttta acatttaaaa tgttaattcc
      121 attttaaatg cacagatgtt tttatttcat aagggtttca atgtgcatga atgctgcaat
      181 attcctgtta ccaaagctag tataaataaa aatagataaa cgtggaaatt acttagagtt
      241 tctgtcatta acgtttcctt cctcagttga caacataaat gcgctgctga gaagccagtt
      301 tgcatctgtc aggatcaatt tcccattatg ccagtcatat taattactag tcaattagtt
      361 gatttttatt tttgacatat acatgtgaaa gaccccacct gtaggtttgg caagctagct
      421 tttccattgc atacgttgta tccatatcat aatatgtaca tttatattgg ctcatgtcca
      481 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      541 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      601 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      661 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      721 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      781 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      841 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      901 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      961 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1021 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1081 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc
     1141 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     1201 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg
     1261 ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa
     1321 gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact gattttatgc
     1381 gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
     1441 acgagttcgg aacacccggc cgcaaccctg ggagacgtcc cagggacttc gggggccgtt
     1501 tttgtggccc gacctgagtc caaaaatccc gatcgttttg gactctttgg tgcacccccc
     1561 ttagaggagg gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
     1621 tctgaatttt tgctttcggt ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc
     1681 atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttgggcccgg
     1741 gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga
     1801 tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag
     1861 aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca
     1921 cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct
     1981 acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac
     2041 accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc
     2101 gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca
     2161 tatggccata tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg
     2221 acatgacaag agttactaac agcccctctc tccaagctca cttacaggct ctctacttag
     2281 tccagcacga agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg
     2341 tggtacctca cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga
     2401 acctagaacc tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca
     2461 aagtagacgg catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg
     2521 gtggaccatc ctctagactg ccggatcgcg caattaaccc tcactaaagg gaacaaaagc
     2581 tggagctcca ccgcggtggc ggccgctaac atggctgact acaaggacga cgatgacaag
     2641 tccatggctg ctgatgtcaa atcgtctcta gtcaatgaat cagaaacgaa tcaaaacagc
     2701 tcctccgatt ccgaggcgga aagacggcct ccgcctcgct ccgaaagttt ccgagacaaa
     2761 tcccgggaaa gtttggaaga agcggccaag aggcaagatg gagggctctt taaggggcca
     2821 ccgtatcccg gctacccctt catcatgatc cccgacctga cgagccccta cctccccaac
     2881 ggatcgctct cgcccaccgc ccgaacctat ctccagatga aatggccact gcttgatgtc
     2941 caggcaggga gcctccagag tagacaagcc ctcaaggatg cccggtcccc atcaccggca
     3001 cacattgtct ctaacaaagt gccagtggtg cagcaccctc accatgtcca ccccctcacg
     3061 cctcttatca cgtacagcaa tgaacacttc acgccgggaa acccacctcc acacttacca
     3121 gccgacgtag accccaaaac aggaatccca cggcctccgc accctccaga tatatccccg
     3181 tattacccac tatcgcctgg caccgtagga caaatccccc atccgctagg atggttagta
     3241 ccacagcaag gtcaaccagt gtacccaatc acgacaggag gattcagaca cccctacccc
     3301 acagctctga ccgtcaatgc ttccatgtcc aggttccctc cccatatggt cccaccacat
     3361 catacgctac acacgacggg cattccgcat ccggccatag tcacaccaac agtcaaacag
     3421 gaatcgtccc agagtgatgt cggctcactc catagttcaa agcatcagga ctccaaaaag
     3481 gaagaagaaa agaagaagcc ccacataaag aaacctctta atgcattcat gttgtatatg
     3541 aaggaaatga gagcaaaggt cgtagctgag tgcacgttga aagaaagcgc ggccatcaac
     3601 cagatccttg ggcggaggtg gcatgcactg tccagagaag agcaagcgaa atactacgag
     3661 ctggcccgga aggagcgaca gcttcatatg caactgtacc ccggctggtc cgcgcgggat
     3721 aactatggaa agaagaagaa gaggaaaagg gacaagcagc cgggagagac caatgaacac
     3781 agcgaatgtt tcctaaatcc ttgcctttca cttcctccga ttacagacct gagcgctcct
     3841 aagaaatgcc gagcgcgctt tggccttgat caacagaata actggtgcgg cccttgcagg
     3901 agaaaaaaaa agtgcgttcg ctacatacaa ggtgaaggca gctgcctcag cccaccctct
     3961 tcagatggaa gcttactaga ttcgcctccc ccctccccga acctgctagg ctcccctccc
     4021 cgagacgcca agtcacagac tgagcagacc cagcctctgt cgctgtccct gaagcccgac
     4081 cccctggccc acctgtccat gatgcctccg ccacccgccc tcctgctcgc tgaggccacc
     4141 cacaaggcct ccgccctctg tcccaacggg gccctggacc tgcccccagc cgctttgcag
     4201 cctgccgccc cctcctcatc aattgcacag ccgtcgactt cttccttaca ttcccacagc
     4261 tccctggccg ggacccagcc ccagccgctg tcgctcgtca ccaagtcttt agaataggaa
     4321 ttccgccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     4381 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4441 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4501 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4561 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4621 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4681 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     4741 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     4801 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     4861 ggttttcctt tgaaaaacac gatgataata tggttgaaca agatggattg cacgcaggtt
     4921 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4981 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5041 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5101 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5161 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5221 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5281 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5341 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5401 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5461 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5521 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5581 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5641 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgatctagc cgccagcaca
     5701 gtggtcgacg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     5761 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     5821 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     5881 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     5941 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6001 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     6061 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     6121 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     6181 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     6241 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     6301 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     6361 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     6421 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6481 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6541 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6601 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6661 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6721 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6781 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6841 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6901 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6961 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7021 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7081 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7141 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7201 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7261 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcactggt aacaggatta
     7321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7381 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7501 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7621 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7681 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7741 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7801 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7861 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7921 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7981 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8041 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8101 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8161 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8221 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8281 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8341 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8401 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8461 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8521 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8581 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8641 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccag
     8701 ctttg
//
