LOCUS       sequence-471715-        6441 bp    DNA     circular SYN 03-JUN-2025
DEFINITION  Retroviral empty vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  TITLE     Mullighan lab plasmids 2025
REFERENCE   2  (bases 1 to 6441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_MSCV-ires-GFP
FEATURES             Location/Qualifiers
     source          1..6441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..942)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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             1129..1643
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(1745..2458,2459..2461,2462..2464))
                     /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(1790..1811)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2139..2158
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2398..2419
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2476..3028
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2478..3045
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2638..2657)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2867..2884
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(3074..3490)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3090..3106)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3122..3144)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3557..3898
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3962..4478
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4678..4697)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4825..4847)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5041..5060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5160..5182
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5220..5238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5306..5410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5411..5479,5480..6271)
                     /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(5629..5648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttttgtggaa ttgtgagcgg
      961 ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa ctatttcagg
     1021 agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac ttaatattcg
     1081 actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat gaaagacccc
     1141 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
     1201 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg cagtctatcg
     1261 gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     1321 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     1381 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggcggg
     1441 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     1501 atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc tgaacttctc
     1561 tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc tagcttgcca
     1621 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
     1681 tttatcgata agcttggctg caggtcgacg gtatcgataa gcttgataat tcttaattaa
     1741 ttcattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     1801 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
     1861 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     1921 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcgccttga
     1981 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     2041 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     2101 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     2161 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     2221 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     2281 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     2341 tgccgtaggt ggcatcgccc tcgccctcgc cagacacgct gaacttgtgg ccgtttacgt
     2401 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     2461 ccatggttgt ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc
     2521 ggggggccta gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc
     2581 cccagatcag atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat
     2641 acgcttgagg agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg
     2701 ggttgtgccg cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg
     2761 tcgccaggtg gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa
     2821 gaagcttcca gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg
     2881 agaggggaaa gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc
     2941 acattgccaa aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct
     3001 tattccaagc ggcttcggcc agtaacgtta gggggggggg gcggaattcg ttaacctcga
     3061 gagatctaat tccggcgcct agagaaggag tgagggctgg ataaagggag gatcgaggcg
     3121 gggtcgaacg aggaggttca agggggagag acggggcgga tggaggaaga ggaggcggag
     3181 gcttagggtg tacaaagggc ttgacccagg gaggggggtc aaaagccaag gcttcccagg
     3241 tcacgatgta ggggacctgg tctgggtgtc catgcgggcc aggtgaaaag accttgatct
     3301 taacctgggt gatgaggtct cggttaaagg tgccgtctcg cggccatccg acgttaaagg
     3361 ttggccattc tgcagagcag aaggtaaccc aacgtctctt cttgacatct accgactggt
     3421 tgtgagcgat ccgctcgaca tctttccagt gacctaaggt caaacttaag ggagtggtaa
     3481 cagtctggcc ctaattttca gacaaataca gaaacacagt cagacagaga caacacagaa
     3541 cgatgctgca gcgctgcagc agacaagacg cgcggcttcg gttccaaacc gaaagcaaaa
     3601 attcagacgg aggcgggaac tgttttaggt tctcgtctcc taccagaacc acatatcctg
     3661 acggggtcgg attccacatc gactcccttc ctcaggtcgg gccacaaaaa cggcccccaa
     3721 agtccctggg acgtctccca gggttgcggc cgggtgttca gaactcgtca gttccaccac
     3781 gggtccgcca gatacagagc tagttagcta actagtacag acgcaggcgc aaacattaga
     3841 tgccggcaca aacacgcaca aagacagaga cagacacgaa acgaccgctg gccagcttac
     3901 ctcccggcgg gggggtcggt ggtccctggg caggggtctc caaatctcgg tggaacctcc
     3961 aaatgaaaga cccccgaggt gggcagtcaa tcaatctgag gagaccctcc caaggatcag
     4021 cgagtccacg attcggatgc aaacagcaag aggctttatt gggaatacgg gtacccgggc
     4081 gacgcagtct atcggaggac tggcgcgccg agtgaggggt tgtgggctct tttattgagc
     4141 tcggggagca gaagcgcgcg aacagaagcg agaagcgaac tgattggtta gttcaaataa
     4201 ggcacagggt catttcaggt ccttggggca ccctggaaac atctgatggt tctctagaaa
     4261 ctgctgaggg cgggaccgca tctggggacc atctgttctt ggccctgagc cggggcagga
     4321 actgcttacc acagatatcc tgtttggccc atattctgct gtctctctgt tcctaacctt
     4381 gatctgaact tctctattct cagttatgta ttttccatgc cttgcaaaat ggcgttactt
     4441 aagctagctt gccaaaccta caggtggggt ctttcattcc cccctttttc tggagactaa
     4501 ataaaatctt ttattttatc tatggctcgt actctatagg cttcagctgg tgatattgtt
     4561 gagtcaaaac tagagcctgg accactgata tcctgtcttt aacaaattgg actaatcgcc
     4621 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     4681 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     4741 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     4801 tgcatgcacc attccttgcg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     4861 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     4921 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     4981 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     5041 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5101 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5161 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5221 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5281 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5341 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5401 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5521 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5581 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5641 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5701 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5821 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5941 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6001 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6241 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6301 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6361 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6421 agaaaagatc aaaggatctt c
//
