LOCUS       sequence-354491-        7627 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 7627)
  AUTHORS   Olive V, Bennett MJ, Walker JC, Ma C, Jiang I, Cordon-Cardo C, Li 
            QJ, Lowe SW, Hannon GJ, He L
  TITLE     miR-19 is a key oncogenic component of mir-17-92.
  JOURNAL   Genes Dev. 2009 Dec 15. 23(24):2839-49.
  PUBMED    20008935
REFERENCE   2  (bases 1 to 7627)
  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: "Genes Dev. 
            2009 Dec 15. 23(24):2839-49."
COMMENT     Sequence Label: MSCV
FEATURES             Location/Qualifiers
     source          1..7627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(38..537)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(86..105)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     479..502
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(572..988)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(588..604)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(620..642)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    1055..1396
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             1460..1976
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2176..2195)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2323..2345)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2539..2558)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2658..2680
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2718..2736)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2804..2908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2909..2977,2978..3769)
                     /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(3127..3146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3940..4528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4429..4448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4682..4699
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4816..4837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4852..4857,4858..4875,4876..4882)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4890..4906
                     /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)."
     LTR             5068..5582
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(5684..6397,6398..6400,6401..6403))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5729..5750)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6078..6097
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6337..6358
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    6415..6967
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6417..6984
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6577..6596)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6806..6823
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(7045..7627,1..17))
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7128..7148)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     join(7625..7627,1..17)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ggcttgtact cggtcatggt aagcttgggc tgcaggtcga aaggcccgga gatgaggaag
       61 aggagaacag cgcggcagac gtgcgctttt gaagcgtgca gaatgccggg cctccggagg
      121 accttcgggc gcccgccccg cccctgagcc cgcccctgag cccgcccccg gacccacccc
      181 ttcccagcct ctgagcccag aaagcgaagg agcaaagctg ctattggccg ctgccccaaa
      241 ggcctacccg cttccattgc tcagcggtgc tgtccatctg cacgagacta gtgagacgtg
      301 ctacttccat ttgtcacgtc ctgcacgacg cgagctgcgg ggcggggggg aacttcctga
      361 ctaggggagg agtagaaggt ggcgcgaagg ggccaccaaa gaacggagcc ggttggcgcc
      421 taccggtgga tgtggaatgt gtgcgaggcc agaggccact tgtgtagcgc caagtgccca
      481 gcggggctgc taaagcgcat gctccagact gccttgggaa aagcgcctcc cctacccggt
      541 agaattcgtt aacctcgaga gatctaattc cggcgcctag agaaggagtg agggctggat
      601 aaagggagga tcgaggcggg gtcgaacgag gaggttcaag ggggagagac ggggcggatg
      661 gaggaagagg aggcggaggc ttagggtgta caaagggctt gacccaggga ggggggtcaa
      721 aagccaaggc ttcccaggtc acgatgtagg ggacctggtc tgggtgtcca tgcgggccag
      781 gtgaaaagac cttgatctta acctgggtga tgaggtctcg gttaaaggtg ccgtctcgcg
      841 gccatccgac gttaaaggtt ggccattctg cagagcagaa ggtaacccaa cgtctcttct
      901 tgacatctac cgactggttg tgagcgatcc gctcgacatc tttccagtga cctaaggtca
      961 aacttaaggg agtggtaaca gtctggccct aattttcaga caaatacaga aacacagtca
     1021 gacagagaca acacagaacg atgctgcagc gctgcagcag acaagacgcg cggcttcggt
     1081 tccaaaccga aagcaaaaat tcagacggag gcgggaactg ttttaggttc tcgtctccta
     1141 ccagaaccac atatcctgac ggggtcggat tccacatcga ctcccttcct caggtcgggc
     1201 cacaaaaacg gcccccaaag tccctgggac gtctcccagg gttgcggccg ggtgttcaga
     1261 actcgtcagt tccaccacgg gtccgccaga tacagagcta gttagctaac tagtacagac
     1321 gcaggcgcaa acattagatg ccggcacaaa cacgcacaaa gacagagaca gacacgaaac
     1381 gaccgctggc cagcttacct cccggcgggg gggtcggtgg tccctgggca ggggtctcca
     1441 aatctcggtg gaacctccaa atgaaagacc cccgaggtgg gcagtcaatc aatctgagga
     1501 gaccctccca aggatcagcg agtccacgat tcggatgcaa acagcaagag gctttattgg
     1561 gaatacgggt acccgggcga cgcagtctat cggaggactg gcgcgccgag tgaggggttg
     1621 tgggctcttt tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg
     1681 attggttagt tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat
     1741 ctgatggttc tctagaaact gctgagggcg ggaccgcatc tggggaccat ctgttcttgg
     1801 ccctgagccg gggcaggaac tgcttaccac agatatcctg tttggcccat attctgctgt
     1861 ctctctgttc ctaaccttga tctgaacttc tctattctca gttatgtatt ttccatgcct
     1921 tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc
     1981 cctttttctg gagactaaat aaaatctttt attttatcta tggctcgtac tctataggct
     2041 tcagctggtg atattgttga gtcaaaacta gagcctggac cactgatatc ctgtctttaa
     2101 caaattggac taatcgcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     2161 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     2221 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     2281 tgttgggcgc catctccttg catgcaccat tccttgcgcc gtcgttttac aacgtcgtga
     2341 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2401 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2461 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2521 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2581 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2641 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     2701 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     2761 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2821 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2881 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2941 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3001 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3061 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3121 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3181 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3241 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3301 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3361 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3421 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3481 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3541 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3601 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3661 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3721 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3781 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3841 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3901 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3961 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4021 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4081 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4141 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4201 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4261 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4321 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4381 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4441 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4501 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4561 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4621 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4681 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4741 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4801 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4861 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acaattctcc
     4921 ccagctcttt ctgcgctgtc cataatgaac tatttcagga gtattggagt atgtggattc
     4981 aaaacagtgg ctaattgagt caggattact taatattcga ctcctaccct cagaatgttg
     5041 ttctccaact tcaagaaact gttacccatg aaagaccccc gctgacgggt agtcaatcac
     5101 tcagaggaga ccctcccaag gaacagcgag accacaagtc ggatgcaact gcaagagggt
     5161 ttattggata cacgggtacc cgggcgacgc agtctatcgg aggactggcg cgccgagtga
     5221 ggggttgtgg gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag
     5281 cgaactgatt ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg
     5341 gaaacatctg atggttctct agaaactgct gagggcggga ccgcatctgg ggaccatctg
     5401 ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt
     5461 ctgctgtctc tctgttccta accttgatct gaacttctct attctcagtt atgtattttc
     5521 catgccttgc aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc
     5581 attcccccct ttttctggag actaaataaa atcttttatt ttatcgataa gcttggctgc
     5641 aggtcgacgg tatcgataag cttgataatt cttaattaat tcattacttg tacagctcgt
     5701 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     5761 tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca
     5821 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc
     5881 cgtcctcgat gttgtggcgg atcttgaagt tcgccttgat gccgttcttc tgcttgtcgg
     5941 ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     6001 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     6061 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     6121 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     6181 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     6241 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     6301 cgccctcgcc agacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     6361 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggttgtg gccatattat
     6421 catcgtgttt ttcaaaggaa aaccacgtcc ccgtggttcg gggggcctag acgttttttt
     6481 aacctcgact aaacacatgt aaagcatgtg caccgaggcc ccagatcaga tcccatacaa
     6541 tggggtacct tctgggcatc cttcagcccc ttgttgaata cgcttgagga gagccatttg
     6601 actctttcca caactatcca actcacaacg tggcactggg gttgtgccgc ctttgcaggt
     6661 gtatcttata cacgtggctt ttggccgcag aggcacctgt cgccaggtgg ggggttccgc
     6721 tgcctgcaaa gggtcgctac agacgttgtt tgtcttcaag aagcttccag aggaactgct
     6781 tccttcacga cattcaacag accttgcatt cctttggcga gaggggaaag acccctagga
     6841 atgctcgtca agaagacagg gccaggtttc cgggccctca cattgccaaa agacggcaat
     6901 atggtggaaa ataacatata gacaaacgca caccggcctt attccaagcg gcttcggcca
     6961 gtaacgttag gggggggggg cggaattgga tgcatggggt cgtgcgctcc tttcggtcgg
     7021 gcgctgcggg tcgtggggcg ggcgtcaggc accgggcttg cgggtcatgc accaggtgcg
     7081 cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg
     7141 gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc
     7201 cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc
     7261 gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat
     7321 ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc
     7381 gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc
     7441 gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag
     7501 ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc
     7561 ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg
     7621 caccgtg
//
