LOCUS       sequence-189206-        6516 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 6516)
  TITLE     Sherr/Roussel lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6516)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMSCV-pBabeMCS-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(592..611)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(739..761)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(955..974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1074..1096
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1134..1152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1220..1324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1325..1393,1394..2185)
                     /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(1543..1562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2356..2944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2845..2864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3098..3115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3232..3253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3268..3273,3274..3291,3292..3298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3306..3322
                     /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             3484..3998
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(4160..4873,4874..4876,4877..4879))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4163..4873,4874..4876,4877..4879))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4205..4226)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4554..4573
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4813..4834
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4882..5431
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5041..5060)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5270..5287
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(5504..5920)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5520..5536)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5552..5574)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5987..6328
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             join(6392..6516,1..392)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt gagctcgggg
       61 agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa ataaggcaca
      121 gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta gaaactgctg
      181 agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc aggaactgct
      241 taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa ccttgatctg
      301 aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt acttaagcta
      361 gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa
      421 tcttttattt tatctatggc tcgtactcta taggcttcag ctggtgatat tgttgagtca
      481 aaactagagc ctggaccact gatatcctgt ctttaacaaa ttggactaat cgcctctacg
      541 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
      601 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
      661 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
      721 caccattcct tgcgccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      781 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      841 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
      901 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
      961 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     1021 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     1081 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     1141 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1561 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1861 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2101 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2161 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3061 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     3241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     3301 tggaattgtg agcggataac aatttcacaa ttctccccag ctctttctgc gctgtccata
     3361 atgaactatt tcaggagtat tggagtatgt ggattcaaaa cagtggctaa ttgagtcagg
     3421 attacttaat attcgactcc taccctcaga atgttgttct ccaacttcaa gaaactgtta
     3481 cccatgaaag acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac
     3541 agcgagacca caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg
     3601 cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag
     3661 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata
     3721 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa
     3781 actgctgagg gcgggaccgc atctggggac catctgttct tggccctgag ccggggcagg
     3841 aactgcttac cacagatatc ctgtttggcc catattctgc tgtctctctg ttcctaacct
     3901 tgatctgaac ttctctattc tcagttatgt attttccatg ccttgcaaaa tggcgttact
     3961 taagctagct tgccaaacct acaggtgggg tctttcattc cccccttttt ctggagacta
     4021 aataaaatct tttattttat cgatatcagc gctttaaatt tgcgcatgct agctatagtt
     4081 ctagaggtac cggttgttaa cgttagccgg ctacctgaag cttgcatgcc tgcaggtcga
     4141 ctctagagtc gcggccgctt tacttgtaca gctcgtccat gccgagagtg atcccggcgg
     4201 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     4261 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     4321 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     4381 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     4441 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     4501 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     4561 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     4621 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     4681 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     4741 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     4801 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     4861 cctcgccctt gctcaccatg gtattatcgt gtttttcaaa ggaaaaccac gtccccgtgg
     4921 ttcggggggc ctagacgttt ttttaacctc gactaaacac atgtaaagca tgtgcaccga
     4981 ggccccagat cagatcccat acaatggggt accttctggg catccttcag ccccttgttg
     5041 aatacgcttg aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac
     5101 tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac
     5161 ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt
     5221 caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg
     5281 gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc
     5341 ctcacattgc caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg
     5401 ccttattcca agcggcttcg gccagtaacg ttaggggggg ggcggaattg atcccgctcg
     5461 agatctgaat tcctacgtac caccacactg ggatcccaat tccggcgcct agagaaggag
     5521 tgagggctgg ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag
     5581 acggggcgga tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg
     5641 gaggggggtc aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc
     5701 catgcgggcc aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg
     5761 tgccgtctcg cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc
     5821 aacgtctctt cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt
     5881 gacctaaggt caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca
     5941 gaaacacagt cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg
     6001 cgcggcttcg gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt
     6061 tctcgtctcc taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc
     6121 ctcaggtcgg gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc
     6181 cgggtgttca gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta
     6241 actagtacag acgcaggcgc aaacattaga tgccggcaca aacacgcaca aagacagaga
     6301 cagacacgaa acgaccgctg gccagcttac ctcccggcgg gggggtcggt ggtccctggg
     6361 caggggtctc caaatctcgg tggaacctcc aaatgaaaga cccccgaggt gggcagtcaa
     6421 tcaatctgag gagaccctcc caaggatcag cgagtccacg attcggatgc aaacagcaag
     6481 aggctttatt gggaatacgg gtacccgggc gacgca
//
