LOCUS       Exported                8792 bp ds-DNA     circular SYN 19-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8792)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 28, 2021 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     Homologous recombination reaction between [pMSCVhyg Vector] as a 
            target sequence  and [PCR(pcDNA5 FRT/TO FLAG-BirAR118G)] as an 
            inserted sequence.
FEATURES             Location/Qualifiers
     source          1..8792
                     /organism="synthetic DNA construct"
                     /mol_type="other 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"
     CDS             1425..1448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1500..2462
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2496..3212
                     /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             3213..3233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3234..3254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        3285..3784
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3802..4827
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             5185..5699
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5861..5877
                     /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(5885..5915)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5930..5951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6239..6827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6998..7858)
                     /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(7859..7963)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(8791..8792,1..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 gatctcccac catggactac aaagacgatg
     1441 acgataaagc cgccagttta aacggtgcgc cattaattaa ggatccagct agcgccacca
     1501 tgaaggacaa caccgtgccc ctgaagctga tcgccctgct ggccaacggc gagttccaca
     1561 gcggcgagca gctgggcgag accctgggca tgagccgggc cgccatcaac aagcacatcc
     1621 agaccctgcg ggactggggc gtggacgtgt tcaccgtgcc cggcaagggc tacagcctgc
     1681 ccgagcccat ccagctgctg aacgccaagc agatcctggg ccagctggac ggcggcagcg
     1741 tggccgtgct gcccgtgatc gacagcacca accagtacct gctggaccgg atcggcgagc
     1801 tgaagagcgg cgacgcctgc atcgccgagt accagcaggc cggccggggc ggccggggcc
     1861 ggaagtggtt cagccccttc ggcgccaacc tgtacctgag catgttctgg cggctggagc
     1921 agggccccgc cgccgccatc ggcctgagcc tggtgatcgg catcgtgatg gccgaggtgc
     1981 tgcggaagct gggcgccgac aaggtgcggg tgaagtggcc caacgacctg tacctgcagg
     2041 accggaagct ggccggcatc ctggtggagc tgaccggcaa gaccggcgac gccgcccaga
     2101 tcgtgatcgg cgccggcatc aacatggcca tgcggcgggt ggaggagagc gtggtgaacc
     2161 agggctggat caccctgcag gaggccggca tcaacctgga ccggaacacc ctggccgcca
     2221 tgctgatccg ggagctgcgg gccgccctgg agctgttcga gcaggagggc ctggccccct
     2281 acctgagccg gtgggagaag ctggacaact tcatcaaccg gcccgtgaag ctgatcatcg
     2341 gcgacaagga gatcttcggc atcagccggg gcatcgacaa gcagggcgcc ctgctgctgg
     2401 agcaggacgg catcatcaag ccctggatgg gcggcgagat cagcctgcgg agcgccgaga
     2461 aggggggatc tggccccggc ggcatggcca caaccatggt gagcaagggc gaggagctgt
     2521 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2581 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2641 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2701 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2761 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2821 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2881 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2941 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3001 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3061 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     3121 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3181 ggatcactct cggcatggac gagctgtaca agcctaagaa gaagaggaag gtgcctaaga
     3241 agaagaggaa ggtgtaagga tcctcgaggt taacgaattc taccgggtag gggaggcgct
     3301 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     3361 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     3421 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     3481 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     3541 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     3601 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     3661 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     3721 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     3781 ttcgacctgc atcccgccac catgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     3841 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     3901 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     3961 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     4021 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     4081 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     4141 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     4201 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     4261 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     4321 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     4381 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     4441 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     4501 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     4561 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     4621 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     4681 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     4741 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     4801 agcactcgtc cgagggcaaa ggaatagagt agatgccgac cgaacaagag ctgatttcga
     4861 gaacgcctca gccagcaact cgcgcgagcc tagcaaggca aatgcgagag aacggcctta
     4921 cgcttggtgg cacagttctc gtccacagtt cgctaagctc gctcggctgg gtcgcgggag
     4981 ggccggtcgc agtgattcag gcccttctgg attgtgttgg tccccagggc acgattgtca
     5041 tgcccacgca ctcgggtgat ctgactgatc ccgcagattg gagatcgccg cccgtgcctg
     5101 ccgattgggt gcagatccgt cgacctgcag ccaagcttat cgataaaata aaagatttta
     5161 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     5221 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     5281 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     5341 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     5401 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     5461 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     5521 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     5581 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     5641 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     5701 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     5761 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     5821 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     5881 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5941 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6001 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6061 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6901 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6961 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7021 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7081 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7141 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7201 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7261 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7321 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7381 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7441 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7501 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7561 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7621 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7681 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7741 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7801 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7861 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7921 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7981 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     8041 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     8101 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     8161 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     8221 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     8281 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     8341 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     8401 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     8461 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     8521 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     8581 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     8641 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     8701 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     8761 gattttattt agtctccaga aaaagggggg aa
//
