LOCUS       Exported                8264 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 8264)
  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..8264
                     /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    1434
                     /label=MCS
                     /note="multiple cloning site"
     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=20 GS
                     /translation="GGSGGSGGSGGGSGGSGGSG"
     CDS             2289..2321
                     /codon_start=1
                     /label=114
                     /translation="VTGYRLFEEIL"
     CDS             2328..3011
                     /codon_start=1
                     /label=CyOFP1
                     /translation="ELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTNRIKVVE
                     GGPLPFAFDILATHFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGGVLTATQ
                     DTSLQDGELIYNVKVRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCDKALKLV
                     GGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARYSNLGGG
                     MDELYK"
     CDS             3021..3074
                     /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             3081..3137
                     /codon_start=1
                     /label=GPA_signal sequence
                     /translation="MYGKIIFVLLLSEIVSISA"
     CDS             3138..3167
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3177..3626
                     /codon_start=1
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLESAGGSAGGSAGGSAGGSAGGPRAQASNSAVDGTAGPGSTGSR"
     CDS             3633..4025
                     /codon_start=1
                     /label=hGPA
                     /translation="LSTTEVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSE
                     ISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPS
                     DVKPLPSPDTDVPLSSVEIENPETSDQ"
     CDS             4035..4094
                     /codon_start=1
                     /label=20GS
                     /translation="GGSGGSGGSGGGSGGSGGSG"
     CDS             4095..4571
                     /codon_start=1
                     /label=11S
                     /translation="VFTLEDFVGDWEQTAAYNLDQVLEQGGVSSLLQNLAVSVTPIQRI
                     VRSGENALKIDIHVIIPYEGLSADQMAQIEEVFKVVYPVDDHHFKVILPYGTLVIDGVT
                     PNMLNYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLITPDGSMLFRVTINS"
     LTR             4626..5140
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5309..5325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5333..5349
                     /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(5357..5387)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5402..5423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5711..6299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6470..7330)
                     /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(7331..7435)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(8263..8264,1..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 cggcagcggc ggctccggtg gatctggcgg tggaagcgga ggttctggtg
     2281 gttctggcgt gacaggctac cggctgttcg aggaaatcct gcggcacgag ctgatcaaag
     2341 aaaacatgcg gagcaagctg tacctggaag gcagcgtgaa cggccaccag ttcaagtgta
     2401 cacacgaagg cgagggcaag ccctacgaag gcaagcagac caaccggatt aaggtggtgg
     2461 aaggcggccc tctgcctttc gcctttgata tcctggccac acactttatg tacggcagca
     2521 aggtgttcat caagtacccc gccgacctgc ctgactactt caagcagagc ttccccgagg
     2581 gcttcacctg ggagcgagtg atggtgtttg aggacggcgg agtgctgacc gccacacagg
     2641 atacttctct gcaggacggc gagctgatct acaacgtgaa agtgcggggc gtgaacttcc
     2701 ccgccaatgg acctgtgatg cagaagaaaa ccctcggctg ggagcccagc accgagacaa
     2761 tgtatccagc cgatggcggc ctcgagggca gatgtgataa ggccctgaaa ctcgtcggcg
     2821 gaggccatct gcacgtgaac tttaagacca cctacaagag caagaaaccc gtgaagatgc
     2881 ccggcgtgca ctacgtggac agacggctgg aaagaatcaa agaggccgac aacgagacat
     2941 acgtggaaca gtacgagcac gccgtggcca gatacagcaa tctcggaggc ggaatggacg
     3001 agctgtacaa aggttctggc gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     3061 agaatcctgg cccaggcgcc atgtatggaa aaataatctt tgtattacta ttgtcagaaa
     3121 ttgtgagcat atcagcagag cagaaactca tctcagaaga ggatctgggc gcgcctatgg
     3181 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     3241 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg
     3301 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     3361 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     3421 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     3481 tggagcttct aaaactggaa agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg
     3541 ctggtggtag tgctggtggt cctcgagctc aagcttcgaa ttctgcagtc gacggtaccg
     3601 cgggcccggg atccaccgga tctagaaccg gtttaagtac cactgaggtg gcaatgcaca
     3661 cttcaacctc ttcttcagtc acaaagagtt acatctcatc acagacaaat gatacgcaca
     3721 aacgggacac atatgcagcc actcctagag ctcatgaagt ttcagaaatt tctgttagaa
     3781 ctgtttaccc tccagaagag gaaaccggag aaagggtaca acttgcccat catttctctg
     3841 aaccagagat aacactcatt atttttgggg tgatggctgg tgttattgga acgatcctct
     3901 taatttctta cggtattcgc cgactgataa agaaaagccc atctgatgta aaacctctcc
     3961 cctcacctga cacagacgtg cctttaagtt ctgttgaaat agaaaatcca gagacaagtg
     4021 atcaattaat taaggggggg tctggaggca gtggtgggtc tggtggtggt tctgggggga
     4081 gtgggggttc aggcgtcttc acactcgaag atttcgttgg ggactgggaa cagacagccg
     4141 cctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgctg cagaatctcg
     4201 ccgtgtccgt aactccgatc caaaggattg tccggagcgg tgaaaatgcc ctgaagatcg
     4261 acatccatgt catcatcccg tatgaaggtc tgagcgccga ccaaatggcc cagatcgaag
     4321 aggtgtttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgccctatg
     4381 gcacactggt aatcgacggg gttacgccga acatgctgaa ctatttcgga cggccgtatg
     4441 aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca
     4501 acaaaattat cgacgagcgc ctgatcaccc ccgacggctc catgctgttc cgagtaacca
     4561 tcaacagcta agcggccgca tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     4621 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     4681 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     4741 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4801 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     4861 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     4921 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     4981 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     5041 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     5101 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
     5161 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
     5221 ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaag
     5281 agctggggag aattaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5521 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6121 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6481 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6541 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6601 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6661 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6721 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6781 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6841 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6901 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6961 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7021 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7081 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7141 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7201 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7261 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7321 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7381 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7441 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7501 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     7561 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     7621 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     7681 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     7741 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     7801 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     7861 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     7921 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     7981 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     8041 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     8101 cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt
     8161 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     8221 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaa
//
