LOCUS       Exported                8877 bp ds-DNA     circular SYN 14-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8877)
  AUTHORS   Leonard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8877
                     /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"
     misc_feature    1437..1493
                     /label=GPA_signal sequence
     CDS             1494..1523
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1533..1811
                     /codon_start=1
                     /label=FRB
                     /translation="MLEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKE
                     TSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISKTR"
     CDS             1818..2210
                     /codon_start=1
                     /label=hGPA
                     /translation="LSTTEVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSE
                     ISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGRIGTILLISYGIRRLIKKSPS
                     DVKPLPSPDTDVPLSSVEIENPETSDQ"
     misc_feature    2067..2069
                     /label=V84R
     CDS             2220..2279
                     /codon_start=1
                     /label=20GS
                     /translation="GGSGGSGGSGGGSGGSGGSG"
     CDS             2280..2312
                     /codon_start=1
                     /label=114
                     /translation="VTGYRLFEEIL"
     CDS             2313..2366
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2388..2444
                     /codon_start=1
                     /label=GPA_signal sequence
                     /translation="MYGKIIFVLLLSEIVSISA"
     CDS             2445..2474
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2484..2933
                     /codon_start=1
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLESAGGSAGGSAGGSAGGSAGGPRAQASNSAVDGTAGPGSTGSR"
     CDS             2940..3332
                     /codon_start=1
                     /label=hGPA
                     /translation="LSTTEVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSE
                     ISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPS
                     DVKPLPSPDTDVPLSSVEIENPETSDQ"
     CDS             3342..3401
                     /codon_start=1
                     /label=20GS
                     /translation="GGSGGSGGSGGGSGGSGGSG"
     CDS             3402..3878
                     /codon_start=1
                     /label=11S
                     /translation="VFTLEDFVGDWEQTAAYNLDQVLEQGGVSSLLQNLAVSVTPIQRI
                     VRSGENALKIDIHVIIPYEGLSADQMAQIEEVFKVVYPVDDHHFKVILPYGTLVIDGVT
                     PNMLNYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLITPDGSMLFRVTINS"
     promoter        3889..4398
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4420..5139
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     LTR             5239..5753
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5922..5938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5946..5962
                     /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(5970..6000)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6015..6036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6324..6912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7083..7943)
                     /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(7944..8048)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(8876..8877,1..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 gatctctcga ccttaagctt ggtaccatgt
     1441 atggaaaaat aatctttgta ttactattgt cagaaattgt gagcatatca gcagagcaga
     1501 aactcatctc agaagaggat ctgggcgcgc ctatgctcga gatgtggcat gaaggcctgg
     1561 aagaggcatc tcgtttgtac tttggggaaa ggaacgtgaa aggcatgttt gaggtgctgg
     1621 agcccttgca tgctatgatg gaacggggcc cccagactct gaaggaaaca tcctttaatc
     1681 aggcctatgg tcgagattta atggaggccc aagagtggtg caggaagtac atgaaatcag
     1741 ggaatgtcaa ggacctcctc caagcctggg acctctatta tcatgtgttc cgacgaatct
     1801 caaagactag aaccggttta agtaccactg aggtggcaat gcacacttca acctcttctt
     1861 cagtcacaaa gagttacatc tcatcacaga caaatgatac gcacaaacgg gacacatatg
     1921 cagccactcc tagagctcat gaagtttcag aaatttctgt tagaactgtt taccctccag
     1981 aagaggaaac cggagaaagg gtacaacttg cccatcattt ctctgaacca gagataacac
     2041 tcattatttt tggggtgatg gctggtcgga ttggaacgat cctcttaatt tcttacggta
     2101 ttcgccgact gataaagaaa agcccatctg atgtaaaacc tctcccctca cctgacacag
     2161 acgtgccttt aagttctgtt gaaatagaaa atccagagac aagtgatcaa ttaattaagg
     2221 gggggtctgg aggcagtggt gggtctggtg gtggttctgg ggggagtggg ggttcaggcg
     2281 tgaccggcta ccggctgttc gaggagattc tggagggcag aggaagtctg ctaacatgcg
     2341 gtgacgtcga ggagaatcct ggcccagata tccttaagct tggtaccatg tatggaaaaa
     2401 taatctttgt attactattg tcagaaattg tgagcatatc agcagagcag aaactcatct
     2461 cagaagagga tctgggcgcg cctatgggag tgcaggtgga aaccatctcc ccaggagacg
     2521 ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag
     2581 atggaaagaa atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
     2641 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
     2701 ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc
     2761 caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaagt gctggtggta
     2821 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtcct cgagctcaag
     2881 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agaaccggtt
     2941 taagtaccac tgaggtggca atgcacactt caacctcttc ttcagtcaca aagagttaca
     3001 tctcatcaca gacaaatgat acgcacaaac gggacacata tgcagccact cctagagctc
     3061 atgaagtttc agaaatttct gttagaactg tttaccctcc agaagaggaa accggagaaa
     3121 gggtacaact tgcccatcat ttctctgaac cagagataac actcattatt tttggggtga
     3181 tggctggtgt tattggaacg atcctcttaa tttcttacgg tattcgccga ctgataaaga
     3241 aaagcccatc tgatgtaaaa cctctcccct cacctgacac agacgtgcct ttaagttctg
     3301 ttgaaataga aaatccagag acaagtgatc aattaattaa gggggggtct ggaggcagtg
     3361 gtgggtctgg tggtggttct ggggggagtg ggggttcagg cgtcttcaca ctcgaagatt
     3421 tcgttgggga ctgggaacag acagccgcct acaacctgga ccaagtcctt gaacagggag
     3481 gtgtgtccag tttgctgcag aatctcgccg tgtccgtaac tccgatccaa aggattgtcc
     3541 ggagcggtga aaatgccctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     3601 gcgccgacca aatggcccag atcgaagagg tgtttaaggt ggtgtaccct gtggatgatc
     3661 atcactttaa ggtgatcctg ccctatggca cactggtaat cgacggggtt acgccgaaca
     3721 tgctgaacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     3781 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcacccccg
     3841 acggctccat gctgttccga gtaaccatca acagctaaca attctaccgg gttggggttg
     3901 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     3961 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     4021 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     4081 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     4141 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     4201 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     4261 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     4321 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     4381 accgacctct ctccccaggg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     4441 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     4501 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     4561 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     4621 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4681 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4741 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4801 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4861 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4921 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4981 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     5041 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     5101 gccgggatca ctctcggcat ggacgagctg tacaagtaac gcccgcccca cgacccgcag
     5161 cgcccgaccg aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt
     5221 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     5281 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
     5341 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     5401 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
     5461 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     5521 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     5581 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
     5641 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5701 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac
     5761 agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg
     5821 actcaattag ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg
     5881 gacagcgcag aagagctggg gagaattaat tcgtaatcat ggtcatagct gtttcctgtg
     5941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8041 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8101 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     8161 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     8221 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     8281 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     8341 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     8401 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     8461 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     8521 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     8581 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     8641 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     8701 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca
     8761 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag
     8821 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaa
//
