LOCUS       sequence-445538-        7799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone of MSCV-driven retroviral cDNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7799)
  AUTHORS   Roe JS, Hwang CI, Somerville TDD, Milazzo JP, Lee EJ, Da Silva B, 
            Maiorino L, Tiriac H, Young CM, Miyabayashi K, Filippini D, 
            Creighton B, Burkhart RA, Buscaglia JM, Kim EJ, Grem JL, Lazenby AJ,
            Grunkemeyer JA, Hollingsworth MA, Grandgenett PM, Egeblad M, Park Y,
            Tuveson DA, Vakoc CR
  TITLE     Enhancer Reprogramming Promotes Pancreatic Cancer Metastasis.
  JOURNAL   Cell. 2017 Aug 24;170(5):875-888.e20. doi: 
            10.1016/j.cell.2017.07.007. Epub 2017 Jul 27.
  PUBMED    28757253
REFERENCE   2  (bases 1 to 7799)
  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"; doi: 
            "10.1016/j.cell.2017.07.007"; journalName: "Cell"; date: 
            "2017-08-24- 24"; volume: "170"; issue: "5"; pages: "875-888.e20"
COMMENT     Sequence Label: MSCV-Neo-GFP/empty
FEATURES             Location/Qualifiers
     source          1..7799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..501
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(36..59)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     434..453
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             515..1318
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(578..597)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1188..1207
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    1387..1955
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1405..1957
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1549..1566)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1776..1795
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1969..1971,1972..1974,1975..2688)
                     /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(2014..2035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2275..2294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2622..2643
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2755..3269
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3431..3447
                     /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(3455..3461,3462..3479,3480..3485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3500..3521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3638..3655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3809..4397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3889..3908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4568..5359,5360..5428))
                     /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     5191..5210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5429..5533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5601..5619
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5657..5679)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5779..5798
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5992..6014
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6142..6161
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6361..6877
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6941..7282
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7349..7765
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7695..7717
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7733..7749
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
       61 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
      121 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
      181 ggaagttccc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
      241 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
      301 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
      361 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
      421 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
      481 tcctcatctc cgggcctttc gacctgcagc caatatggga tcggccattg aacaagatgg
      541 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      601 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
      661 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
      721 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
      781 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
      841 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
      901 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
      961 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     1021 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     1081 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     1141 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     1201 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     1261 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagg
     1321 ggatcccgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
     1381 tccaattccg cccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     1441 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     1501 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     1561 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     1621 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     1681 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     1741 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     1801 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     1861 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     1921 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     1981 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2041 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     2101 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2161 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2221 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2281 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2341 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2401 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     2461 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2521 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2581 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2641 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacctgcag
     2701 ccaagcttat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     2761 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     2821 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
     2881 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     2941 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
     3001 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     3061 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     3121 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc
     3181 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     3241 ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac
     3301 attctgaggg taggagtcga atattaagta atcctgactc aattagccac tgttttgaat
     3361 ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag
     3421 aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3481 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3541 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3601 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3661 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3721 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3781 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3841 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3901 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3961 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4021 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4081 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4141 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4201 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4261 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4321 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4561 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4621 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4681 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4741 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4801 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4861 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4921 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4981 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5041 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5101 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5161 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5221 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5281 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5341 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5401 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5521 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5581 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     5641 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5701 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5761 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     5821 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     5881 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     5941 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     6001 gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     6061 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6121 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6181 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa
     6241 agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc
     6301 tatagagtac gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg
     6361 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     6421 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
     6481 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     6541 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
     6601 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     6661 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     6721 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
     6781 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
     6841 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
     6901 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
     6961 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
     7021 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
     7081 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
     7141 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
     7201 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
     7261 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
     7321 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     7381 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     7441 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     7501 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     7561 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     7621 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     7681 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     7741 agccctcact ccttctctag gcgccggaat tcgttaacct cgagagatct aattctacc
//
