LOCUS       Exported                7384 bp ds-DNA     circular UNA 30-JUL-2023
DEFINITION  natural circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMSCV-FLAG-p-mOct4 S12A
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7384)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 31, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7384
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     regulatory      1416..1425
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1425..1448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1458..2516
                     /codon_start=1
                     /label=mOct4
                     /translation="MAGHLASDFAFAPPPGGGDGSAGLEPGWVDPRTWLSFQGPPGGPG
                     IGPGSEVLGISPCPPAYEFCGGMAYCGPQVGLGLVPQVGVETLQPEGQAGARVESNSEG
                     TSSEPCADRPNAVKLEKVEPTPEESQDMKALQKELEQFAKLLKQKRITLGYTQADVGLT
                     LGVLFGKVFSQTTICRFEALQLSLKNMCKLRPLLEKWVEEADNNENLQEICKSETLVQA
                     RKRKRTSIENRVRWSLETMFLKCPKPSLQQITHIANQLGLEKDVVRVWFCNRRQKGKRS
                     SIEYSQREEYEATGTPFPGGAVSFPLPPGPHFGTPGYGSPHFTTLYSVPFPEGEAFPSV
                     PVTALGSPMHSN"
     misc_feature    1491..1493
                     /label=S12A
     promoter        2527..3026
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3047..3646
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             3746..4260
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4429..4445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4453..4469
                     /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(4477..4507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4522..4543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4831..5419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5590..6450)
                     /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"
     promoter        complement(6451..6555)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(7383..7384,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctgccac catggattac aaggatgacg
     1441 acgataagct cgagaccatg gctggacacc tggcttcaga cttcgccttc gcacccccac
     1501 caggtggggg tgatgggtca gcagggctgg agccgggctg ggtggatcct cgaacctggc
     1561 taagcttcca agggcctcca ggtgggcctg gaatcggacc aggctcagag gtattgggga
     1621 tctccccatg tccgcccgca tacgagttct gcggagggat ggcatactgt ggacctcagg
     1681 ttggactggg cctagtcccc caagttggcg tggagacttt gcagcctgag ggccaggcag
     1741 gagcacgagt ggaaagcaac tcagagggaa cctcctctga gccctgtgcc gaccgcccca
     1801 atgccgtgaa gttggagaag gtggaaccaa ctcccgagga gtcccaggac atgaaagccc
     1861 tgcagaagga gctagaacag tttgccaagc tgctgaagca gaagaggatc accttggggt
     1921 acacccaggc cgacgtgggg ctcaccctgg gcgttctctt tggaaaggtg ttcagccaga
     1981 ccaccatctg tcgcttcgag gccttgcagc tcagccttaa gaacatgtgt aagctgcggc
     2041 ccctgctgga gaagtgggtg gaggaagccg acaacaatga gaaccttcag gagatatgca
     2101 aatcggagac cctggtgcag gcccggaaga gaaagcgaac tagcattgag aaccgtgtga
     2161 ggtggagtct ggagaccatg tttctgaagt gcccgaagcc ctccctacag cagatcactc
     2221 acatcgccaa tcagcttggg ctagagaagg atgtggttcg agtatggttc tgtaaccggc
     2281 gccagaaggg caaaagatca agtattgagt attcccaacg agaagagtat gaggctacag
     2341 ggacaccttt cccagggggg gctgtatcct ttcctctgcc cccaggtccc cactttggca
     2401 ccccaggcta tggaagcccc cacttcacca cactctactc agtccctttt cctgagggcg
     2461 aggcctttcc ctctgttccc gtcactgctc tgggctctcc catgcattca aactgagaat
     2521 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     2581 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     2641 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     2701 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     2761 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     2821 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     2881 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     2941 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     3001 cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga ccgagtacaa
     3061 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3121 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3181 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3241 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3301 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3361 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3421 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3481 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3541 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3601 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     3661 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt
     3721 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     3781 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     3841 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     3901 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     3961 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4021 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4081 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     4141 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4201 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     4261 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     4321 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     4381 cattatggac agcgcagaag agctggggag aattaattcg taatcatggt catagctgtt
     4441 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4501 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4561 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4621 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4681 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4741 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4801 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4861 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4921 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4981 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5041 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5101 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5161 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5221 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5281 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5341 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5401 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5461 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5581 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5641 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5701 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5761 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5821 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5881 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5941 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6001 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6061 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6121 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6181 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6241 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6301 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6361 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6421 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6481 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6541 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6601 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6661 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6721 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6781 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6841 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6901 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6961 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7021 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     7081 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     7141 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     7201 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     7261 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     7321 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     7381 ggaa
//
