LOCUS       Exported                7181 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 7181)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7181
                     /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    1446..1502
                     /label=GPA_signal sequence
     CDS             1503..1532
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1542..1820
                     /codon_start=1
                     /label=FRB
                     /translation="MLEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKE
                     TSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISKTR"
     CDS             1827..2219
                     /codon_start=1
                     /label=hGPA
                     /translation="LSTTEVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSE
                     ISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPS
                     DVKPLPSPDTDVPLSSVEIENPETSDQ"
     CDS             2229..2288
                     /codon_start=1
                     /label=20GS
                     /translation="GGSGGSGGSGGGSGGSGGSG"
     CDS             2289..2795
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     CERILA"
     CDS             2802..3485
                     /codon_start=1
                     /label=CyOFP1
                     /translation="ELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTNRIKVVE
                     GGPLPFAFDILATHFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGGVLTATQ
                     DTSLQDGELIYNVKVRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCDKALKLV
                     GGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARYSNLGGG
                     MDELYK"
     LTR             3543..4057
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4226..4242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4250..4266
                     /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(4274..4304)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4319..4340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4628..5216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5387..6247)
                     /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(6248..6352)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(7180..7181,1..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 ggttaacgaa ttcgctagcg
     1441 ccaccatgta tggaaaaata atctttgtat tactattgtc agaaattgtg agcatatcag
     1501 cagagcagaa actcatctca gaagaggatc tgggcgcgcc tatgctcgag atgtggcatg
     1561 aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa ggcatgtttg
     1621 aggtgctgga gcccttgcat gctatgatgg aacggggccc ccagactctg aaggaaacat
     1681 cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc aggaagtaca
     1741 tgaaatcagg gaatgtcaag gacctcctcc aagcctggga cctctattat catgtgttcc
     1801 gacgaatctc aaagactaga accggtttaa gtaccactga ggtggcaatg cacacttcaa
     1861 cctcttcttc agtcacaaag agttacatct catcacagac aaatgatacg cacaaacggg
     1921 acacatatgc agccactcct agagctcatg aagtttcaga aatttctgtt agaactgttt
     1981 accctccaga agaggaaacc ggagaaaggg tacaacttgc ccatcatttc tctgaaccag
     2041 agataacact cattattttt ggggtgatgg ctggtgttat tggaacgatc ctcttaattt
     2101 cttacggtat tcgccgactg ataaagaaaa gcccatctga tgtaaaacct ctcccctcac
     2161 ctgacacaga cgtgccttta agttctgttg aaatagaaaa tccagagaca agtgatcaat
     2221 taattaaggg ggggtctgga ggcagtggtg ggtctggtgg tggttctggg gggagtgggg
     2281 gttcaggctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     2341 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     2401 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     2461 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     2521 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     2581 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     2641 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     2701 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     2761 tgaccggctg gcggctgtgc gaacgcattc tggcgcggca cgagctgatc aaagaaaaca
     2821 tgcggagcaa gctgtacctg gaaggcagcg tgaacggcca ccagttcaag tgtacacacg
     2881 aaggcgaggg caagccctac gaaggcaagc agaccaaccg gattaaggtg gtggaaggcg
     2941 gccctctgcc tttcgccttt gatatcctgg ccacacactt tatgtacggc agcaaggtgt
     3001 tcatcaagta ccccgccgac ctgcctgact acttcaagca gagcttcccc gagggcttca
     3061 cctgggagcg agtgatggtg tttgaggacg gcggagtgct gaccgccaca caggatactt
     3121 ctctgcagga cggcgagctg atctacaacg tgaaagtgcg gggcgtgaac ttccccgcca
     3181 atggacctgt gatgcagaag aaaaccctcg gctgggagcc cagcaccgag acaatgtatc
     3241 cagccgatgg cggcctcgag ggcagatgtg ataaggccct gaaactcgtc ggcggaggcc
     3301 atctgcacgt gaactttaag accacctaca agagcaagaa acccgtgaag atgcccggcg
     3361 tgcactacgt ggacagacgg ctggaaagaa tcaaagaggc cgacaacgag acatacgtgg
     3421 aacagtacga gcacgccgtg gccagataca gcaatctcgg aggcggaatg gacgagctgt
     3481 acaaatgagc ggccgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3541 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     3601 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     3661 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3721 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     3781 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3841 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3901 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     3961 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     4021 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     4081 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     4141 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaagagc
     4201 tggggagaat taattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4261 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4321 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4381 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4801 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5041 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6361 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6421 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6481 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6541 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6601 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6661 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     6721 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6781 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc
     6841 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     6901 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     6961 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     7021 gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca
     7081 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
     7141 aaaataaaag attttattta gtctccagaa aaagggggga a
//
