LOCUS       sequence-221993-        5551 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 5551)
  AUTHORS   Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz 
            L, Nusse R, Weissman IL
  TITLE     A role for Wnt signalling in self-renewal of haematopoietic stem 
            cells.
  JOURNAL   Nature. 2003 May 22. 423(6938):409-14.
  PUBMED    12717450
REFERENCE   2  (bases 1 to 5551)
  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: "Nature. 
            2003 May 22. 423(6938):409-14."
COMMENT     Sequence Label: VSV.G
FEATURES             Location/Qualifiers
     source          1..5551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        494..697
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     648..668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     694..718
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          833..965
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     948..967
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1010..1028
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1052..1099,1100..2587)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     polyA_signal    2641..2762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2652..2671)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2706..2725
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2943..3398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3030..3049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3240..3261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3465..3484)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3584..3606
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3644..3662)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3730..3834
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3835..3903,3904..4695)
                     /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     complement(4053..4072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4866..5454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5355..5374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
       61 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttggcattg
      121 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      181 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      241 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      301 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      361 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      421 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
      481 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
      541 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      601 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
      661 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcac
      721 tagaagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca gtgcttctga
      781 cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat
      841 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
      901 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
      961 cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc
     1021 actataggct agcctcgaga attctgacac tatgaagtgc cttttgtact tagccttttt
     1081 attcattggg gtgaattgca agttcaccat agtttttcca cacaaccaaa aaggaaactg
     1141 gaaaaatgtt ccttctaatt accattattg cccgtcaagc tcagatttaa attggcataa
     1201 tgacttaata ggcacagcct tacaagtcaa aatgcccaag agtcacaagg ctattcaagc
     1261 agacggttgg atgtgtcatg cttccaaatg ggtcactact tgtgatttcc gctggtatgg
     1321 accgaagtat ataacacatt ccatccgatc cttcactcca tctgtagaac aatgcaagga
     1381 aagcattgaa caaacgaaac aaggaacttg gctgaatcca ggcttccctc ctcaaagttg
     1441 tggatatgca actgtgacgg atgccgaagc agtgattgtc caggtgactc ctcaccatgt
     1501 gctggttgat gaatacacag gagaatgggt tgattcacag ttcatcaacg gaaaatgcag
     1561 caattacata tgccccactg tccataactc tacaacctgg cattctgact ataaggtcaa
     1621 agggctatgt gattctaacc tcatttccat ggacatcacc ttcttctcag aggacggaga
     1681 gctatcatcc ctgggaaagg agggcacagg gttcagaagt aactactttg cttatgaaac
     1741 tggaggcaag gcctgcaaaa tgcaatactg caagcattgg ggagtcagac tcccatcagg
     1801 tgtctggttc gagatggctg ataaggatct ctttgctgca gccagattcc ctgaatgccc
     1861 agaagggtca agtatctctg ctccatctca gacctcagtg gatgtaagtc taattcagga
     1921 cgttgagagg atcttggatt attccctctg ccaagaaacc tggagcaaaa tcagagcggg
     1981 tcttccaatc tctccagtgg atctcagcta tcttgctcct aaaaacccag gaaccggtcc
     2041 tgctttcacc ataatcaatg gtaccctaaa atactttgag accagataca tcagagtcga
     2101 tattgctgct ccaatcctct caagaatggt cggaatgatc agtggaacta ccacagaaag
     2161 ggaactgtgg gatgactggg caccatatga agacgtggaa attggaccca atggagttct
     2221 gaggaccagt tcaggatata agtttccttt atacatgatt ggacatggta tgttggactc
     2281 cgatcttcat cttagctcaa aggctcaggt gttcgaacat cctcacattc aagacgctgc
     2341 ttcgcaactt cctgatgatg agagtttatt ttttggtgat actgggctat ccaaaaatcc
     2401 aatcgagctt gtagaaggtt ggttcagtag ttggaaaagc tctattgcct cttttttctt
     2461 tatcataggg ttaatcattg gactattctt ggttctccga gttggtatcc atctttgcat
     2521 taaattaaag cacaccaaga aaagacagat ttatacagac atagagatga accgacttgg
     2581 aaagtaactc aaatcctgca ctctagagtc gacccgggcg gccgcttcga gcagacatga
     2641 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2701 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2761 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2821 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg ggctggcgta
     2881 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2941 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3001 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3061 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3121 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3181 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3241 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3301 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3361 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     3421 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     3481 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3541 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3601 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3661 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3721 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3781 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3841 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3901 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3961 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4021 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4081 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4141 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4201 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4261 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4321 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4381 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4441 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4501 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4561 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4621 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4681 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4741 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4801 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4981 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5341 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5401 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5461 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     5521 cagatcttca atattggcca ttagccatat t
//
