LOCUS       Exported                6330 bp DNA     circular SYN 17-OCT-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Oct 17, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6330
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        25..265
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    25..265
                     /label=U6 promoter
     misc_feature    274..301
                     /label=U6+27
     misc_feature    308..318
                     /label=P3
     misc_feature    318..372
                     /label=TTRz
                     /note="Original from Nat Chem Bio Lilley Structure 2014:"
     misc_feature    join(319..321,332..334)
                     /label=P4
     misc_feature    340..343
                     /label=P2
     misc_feature    join(349..353,358..362)
                     /label=P1
     misc_feature    354..357
                     /label=tetraloop
     misc_feature    362..377
                     /label=Clonado-F30-F1
     misc_feature    364..369
                     /label=5'tRNA
     misc_feature    364
                     /label=Req AA
     misc_feature    364
                     /label=Post Cleave
     misc_feature    366..369
                     /label=P2
     misc_feature    370..374
                     /label=Spacer
     misc_feature    372..377
                     /label=P3
     misc_feature    385..401
                     /label=F30_5'
     misc_feature    402..425
                     /label=SAM5'
     misc_feature    430..492
                     /label=Squash_0bp
     misc_feature    497..507
                     /label=SAM3'
     misc_feature    508..525
                     /label=F30-Mid
     misc_feature    526..574
                     /label=Broccoli
     misc_feature    575..589
                     /label=F30-3'
     misc_feature    600..604
                     /label=anti-Spacer
     misc_feature    603..614
                     /label=anti-P1 CleaveStem
                     /note="Original from Nat Chem Bio Lilley Structure 2014:
                     5' CCGCC T
                     
                     GGCGG 3'
                     "
     misc_feature    605..615
                     /label=3'tRNA
                     /label=intron(1)
     misc_feature    605..615
                     /label=P1 CleaveStem
                     /note="Original from Nat Chem Bio Lilley Structure 2014:
                     5' CCGCC T
                     
                     GGCGG 3'
                     "
     misc_feature    616..658
                     /label=TRz
                     /note="Original from Nat Chem Bio Lilley Structure 2014:"
     misc_feature    616
                     /label=Post Cleave1
     misc_feature    join(618..621,650..653)
                     /label=P2
     misc_feature    625..627
                     /label=dP3
     misc_feature    join(629..632,641..644)
                     /label=P4
     misc_feature    659..670
                     /label=anti-P1 CleaveStem
                     /note="Original from Nat Chem Bio Lilley Structure 2014:
                     5' CCGCC T
                     
                     GGCGG 3'
                     "
     misc_feature    672..696
                     /label=U6 term
     misc_feature    683..686
                     /label=TL
     polyA_signal    781..902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1027..1455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1482..1586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1587..2447
                     /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"
     polyA_signal    3649..3783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3662..3783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4208..4228)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4358..4423
                     /label=small t intron
                     /note="simian virus 40 (SV40) small t antigen intron"
     promoter        complement(5993..6322)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6007..6142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gccggatcca aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat
       61 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
      121 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      181 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      241 gctttatata tcttgtggaa aggacgaaac accgtgctcg cttcggcagc acatatacta
      301 gtcgacggcc atcagtcgcc ggtcccaagc ccggataaaa tgggaggggg cgggaaaccg
      361 cctaaccatg ccgactgatg gcagttgcca tgtgtatgtg gccagatgcc ttgtaaccga
      421 aagggaatca aggtgagccc aataatacgg tttgggttag gataggaagt agagccgtaa
      481 actctctaag cggattcgaa aggatggcca catactctga tgatcgagac ggtcgggtcc
      541 agatattcgt atctgtcgag tagagtgtgg gctcgatcat tcatggcaac tgccatcagt
      601 cggcgtggac tgtagaacac tgccaatgcc ggtcccaagc ccggataaaa gtggagggta
      661 cagtccacgc tctagagcgc acttcggtgc gctttttact aggacctgca ggcatgcaag
      721 cttgacgtcg gttaccgata tccatatggc gaccgcatcg atctcgagcc gaggactagt
      781 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      841 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      901 tatcatgtct tacgtagata agtagcatgg cgggttatag cgaagaggcc cgcaccgatc
      961 gcccttccca acagttgcgc agcctgaatg gctaatggga aattgtaaac gttaatattt
     1021 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     1081 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     1141 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     1201 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     1261 ggtgccgtaa agcactaaat cggaacccta aagggatgcc ccgatttaga gcttgacggg
     1321 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     1381 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     1441 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1501 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1561 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1621 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1681 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1741 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1801 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1861 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1921 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1981 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2041 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2101 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2161 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2221 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2281 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2341 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2401 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2461 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2521 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2581 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2641 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2701 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2761 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2821 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2881 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2941 ggggttcgtg caacacagcc agcttggagc gaacgaccta caccgaactg agatacctac
     3001 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3061 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3301 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3361 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3421 gttggccgat tcattaatgc agagatcttt ggccactccc tctctgcgcg ctcgctcgct
     3481 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3541 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctggagggg
     3601 tggagtcgtg acgtgaatta cgtcataggg ttagggaggt cctggatcga tccagacatg
     3661 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3721 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3781 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3841 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct ctagtcaagg
     3901 cactatacat caaatattcc ttattaaccc ctttacaaat taaaaagcta aaggtacaca
     3961 atttttgagc atagttatta atagcagaca ctctatgcct gtgtggagta agaaaaaaca
     4021 gtatgttatg attataactg ttatgcctac ttataaaggt tacagaatat ttttccataa
     4081 ttttcttgta tagcagtgca gctttttcct ttgtggtgta aatagcaaag caagcaagag
     4141 ttctattact aaacacagca tgactcaaaa aacttagcaa ttctgaagga aagtccttgg
     4201 ggtcttctac ctttctcttc ttttttggag gagtagaatg ttgagagtca gcagtagcct
     4261 catcatcact agatggcatt tcttctgagc aaaacaggtt ttcctcatta aaggcattcc
     4321 accactgctc ccattcatca gttccatagg ttggaatcta aaatacacaa acaattagaa
     4381 tcagtagttt aacacattat acacttaaaa attttatatt taccttagag ctttaaatct
     4441 ctgtaggtag tttgtccaat tatgtcacac cacagaagta aggttccttc acaaagatcc
     4501 gggaccaaag cggccatcgt gcctccccac tcctgcagtt cgggggcatg gatgcgcgga
     4561 tagccgctgc tggtttcctg gatgccgacg gatttgcact gccggtagaa ctccgcgagg
     4621 tcgtccagcc tcaggcagca gctgaaccaa ctcgcgaggg gatcgagccc ggggtgggcg
     4681 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     4741 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     4801 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     4861 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     4921 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4981 cctgatagcg cgccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     5041 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     5101 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     5161 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     5221 atgttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     5281 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     5341 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     5401 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     5461 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     5521 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     5581 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     5641 aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca gatccttggc
     5701 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     5761 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcggcatgta
     5821 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     5881 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgttc
     5941 cgcttccttt agcagccctt gcgccctgag tgcttgcggc agcgtgaagc tttttgcaaa
     6001 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     6061 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     6121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     6181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     6241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     6301 ttccacaccc taactgacac acattccaca
//
