LOCUS       sequence-322907-        8339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral coexpression of Cre and EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8339)
  TITLE     Littman Lab Plasmids
REFERENCE   2  (bases 1 to 8339)
  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"
COMMENT     Sequence Label: MSCV-Cre
FEATURES             Location/Qualifiers
     source          1..8339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(455..1246,1247..1315))
                     /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     1078..1097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1316..1420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1488..1506
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1544..1566)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1666..1685
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1879..1901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2029..2048
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2248..2764
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2828..3169
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3236..3652
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3582..3604
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3620..3636
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3675..3695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3696..4724
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     intron          5054..5626
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        5685..6184
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5720..5743)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6117..6136
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      6202..6211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6208..6210,6211..6213,6214..6927)
                     /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             join(6208..6210,6211..6213,6214..6924)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6514..6533)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6861..6882
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6981..7495
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7657..7673
                     /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(7681..7687,7688..7705,7706..7711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7726..7747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7864..7881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8035..8339,1..284))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8115..8134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
       61 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1441 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1501 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1561 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1621 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1681 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1741 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1801 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1861 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     1921 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     1981 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2041 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2101 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     2161 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     2221 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     2281 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     2341 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     2401 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     2461 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     2521 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     2581 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     2641 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     2701 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
     2761 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc
     2821 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc
     2881 cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     2941 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     3001 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     3061 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     3121 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     3181 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     3241 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     3301 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     3361 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     3421 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     3481 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     3541 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     3601 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag
     3661 atctctcgac catgcccaag aagaagagga aggtgtccaa tttactgacc gtacaccaaa
     3721 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
     3781 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
     3841 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
     3901 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc
     3961 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
     4021 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
     4081 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
     4141 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
     4201 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
     4261 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
     4321 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
     4381 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     4441 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     4501 gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca
     4561 gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca
     4621 tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa ctatatccgt aacctggata
     4681 gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga ttagccatta acgcgtaaat
     4741 gattgctata attatttgat atttatggtg acatatgaga aaggatttca acatcgacgg
     4801 aaaatatgta gtgctgtctg taagcactaa tattcagtcg ccagccgtca ttgtcactgt
     4861 aaagctgagc gatagaatgc ctgatattga ctcaatatcc gttgcgtttc ctgtcaaaag
     4921 tatgcgtagt gctgaacatt tcgtgatgaa tgccaccgag gaagaagcac ggcgcggttt
     4981 tgctaaagtg atgtctgagt ttggcgaact cttgggtaag gttggaattg tcgatagagc
     5041 tgagaacttc agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa
     5101 attcatgtta tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc
     5161 cttgtatcac catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca
     5221 accattgtct cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt
     5281 gcatttgtaa cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct
     5341 ctaatcactt ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta
     5401 gagaacaatt gttataatta aatgataagg tagaatattt ctgcatataa attctggctg
     5461 gcgtggaaat attcttattg gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc
     5521 tttatggtta caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc
     5581 gggcccctct gctaaccatg ttcatgcctt cttctctttc ctacagctcc tgggcaacgt
     5641 gctggttgtt gtgctgtctc atcattttgg caaagaattc taccgggtag gggaggcgct
     5701 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     5761 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     5821 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     5881 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     5941 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     6001 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     6061 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     6121 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     6181 ttcgacctgc agcccaagct tgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     6241 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     6301 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     6361 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     6421 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     6481 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     6541 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     6601 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     6661 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     6721 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     6781 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     6841 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     6901 ctcggcatgg acgagctgta caagtaaagc ggccatcgat aaaataaaag attttattta
     6961 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     7021 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     7081 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     7141 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     7201 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     7261 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     7321 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     7381 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     7441 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta
     7501 acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc
     7561 tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta
     7621 tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca
     7681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8221 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatc
//
