LOCUS       E8GGMX5_pYAO_CMV        6635 bp    DNA     circular SYN 05-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6635)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1..241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          4242..4268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..241
                     /label=U6 promoter RNA polymerase III promoter for hum
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(231..250)
                     /label=xct228
     primer_bind     252..322
                     /label=441_GMX5_F
     primer_bind     complement(256..326)
                     /label=442_GMX5_R
     misc_feature    256..306
                     /label=X1
     misc_feature    307..322
                     /label=spacer
     misc_feature    323..500
                     /label=wRNA
     primer_bind     complement(472..547)
                     /label=xct113_wRpAsig_F1
     polyA_signal    512..567
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(518..595)
                     /label=xct114_wRpAsig_F2
     primer_bind     564..583
                     /label=xct097_wRpA_GGseq
     primer_bind     complement(578..601)
                     /label=xct109_wRNAgg_F
     polyA_signal    596..820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     598..615
                     /label=xct108_TmCgg_R
     primer_bind     659..668
                     /label=411_GX1_R
     CDS             complement(858..1568)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFK*ERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1251..1317
                     /label=441_GMX5_F
     primer_bind     complement(1251..1317)
                     /label=442_GMX5_R
     primer_bind     1261..1327
                     /label=437_GX3_F
     primer_bind     1265..1332
                     /label=435_GMX2_F
     primer_bind     1271..1337
                     /label=434_GMX1_F
     CDS             complement(1569..1622)
                     /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"
     primer_bind     complement(1604..1653)
                     /label=xct319_E8_F
     CDS             complement(1629..1655)
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
     primer_bind     1634..1686
                     /label=xct318_ADAR_R
     CDS             complement(1668..2822)
                     /codon_start=1
                     /label=ADARdd
                     /translation="QLHLPQVLADAVSRLVLGKFGDLTDNFSSPHARRKVLAGVVMTTG
                     TDVKDAKVISVSTGGKCINGEYMSDRGLALNDCHAEIISRRSLLRFLYTQLELYLNNKD
                     DQKRSIFQKSERGGFRLKENVQFHLYISTSPCGDARIFSPHEPILEEPADRHPNRKARG
                     QLRTKIESGQGTIPVRSNASIQTWDGVLQGERLLTMSCSDKIARWNVVGIQGSLLSIFV
                     EPIYFSSIILGSLYHGDHLSRAMYQRISNIEDLPPLYTLNKPLLSGISNAEARQPGKAP
                     NFSVNWTVGDSAIEVINATTGKDELGRASRLCKHALYCRWMRVHGKVPSHLLRSKITKP
                     NVYHESKLAAKEYQAAKARLFTAFIKAGLGAWVEKPTEQDQFSLT"
     misc_feature    2304..2306
                     /label=E488Q
     misc_feature    2643..2645
                     /label=T375G
     primer_bind     complement(2804..2873)
                     /label=xct317_ADAR_F
     misc_feature    2823..2918
                     /label=32aalinker
     primer_bind     2854..2937
                     /label=xct316_E8_R
     CDS             complement(2919..4241)
                     /codon_start=1
                     /label=IscB
                     /translation="MAVVYVISKSGKPLMPTTRCGHVRILLKEGKARVVERKPFTIQLT
                     YESAEETQPLVLGIAPGRTNIGMSVVTESGESVFNAQIETRNKDVPKLMKDRKQYRRAH
                     RRLKRRCKRRRRAKAAGTAFEEGEKQRLLPGCKKPITCKSIRNKEARFNNRKRPKGWLT
                     PTANHLLVTHLNVVKKVQKILPVAKVVLELNRFSFMAMNNPKVQRWQYQRGPLYGKGSV
                     EEAVSMQQDGHCLFCKHGIDHYHHVVPRRKNGSETLENRVGLCEEHARLVHTDKEWEAN
                     LASKKSGMNKKYHALSVLNQIIPYLADQLADMFPGNFCVTSGQDTYLFREEHGIPKDHY
                     LDAYCIACSALTDAKKVSSPKGRPYMVHQFRRHDRQACHKANLNRSYYMGGKKVATNRH
                     KAMDQKTDSLEEYRAAHSAADVSKLTVKHPSAQYNGGLQIYV"
     primer_bind     complement(2921..2952)
                     /label=xct177_E5dT_F
     primer_bind     2933..2964
                     /label=xct178_E5dT_R
     primer_bind     complement(2945..2964)
                     /label=xct096_IscBC_GGseq
     primer_bind     2956..2966
                     /label=434_GMX1_F
     misc_feature    3063..3065
                     /label=L393K
     primer_bind     complement(3159..3177)
                     /label=xct175_EIscB_seq
     misc_feature    3435..3437
                     /label=H269A
     misc_feature    3765..3767
                     /label=V159K
     misc_feature    3831..3833
                     /label=F137K
     misc_feature    3936..3938
                     /label=M102R
     misc_feature    4062..4064
                     /label=D60A
     primer_bind     complement(4221..4261)
                     /label=xct166_EIscBWT_F
     CDS             complement(4242..4274)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=new SV40 NLS
                     /label=SV40 NLS
                     /translation="MGPKKKRKVGS"
     primer_bind     4242..4260
                     /label=xct165_NNLS_R
     misc_feature    4275..4280
                     /label=Kozak
     misc_feature    4281..4300
                     /label=arm2
     promoter        complement(4305..4508)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4334..4354)
                     /label=xct164_CMV_seq
     enhancer        4509..4888
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4855..4884)
                     /label=xct103_CMVgg_F
     primer_bind     4881..4905
                     /label=xct115_YAObb_R
     promoter        4896..5000
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             join(5001..5069,5070..5861)
                     /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"
     CDS             5001..5112
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQ"
     primer_bind     complement(5005..5026)
                     /label=xct094A_Amp_GGseq
     CDS             5113..5861
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="GARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRID
                     AGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGG
                     PKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLAS
                     RQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIY
                     TTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      6032..6620
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     primer_bind     6457..6475
                     /label=ori-t-R
     primer_bind     complement(6603..6624)
                     /label=xct116_YAObb_F
     primer_bind     join(6621..6635,1..18)
                     /label=xct110_wRNAgg_R
     misc_feature    6621..6635
                     /label=MCS
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gtcctgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa
      301 ggacagcttc aagtagtcgg ggggctcttt caacttgaaa aggttgaaag agcacaggct
      361 gagacattcg taaggccgaa aggccggacg caccctggga tttccccagt ccccggaact
      421 gcatagcgga tgtcagttga tgaaaatcag ataagccagg gggaacaatc acctctctgg
      481 aaacagagag agtttttttt gttttgcagt aaataaagga aatttatttt cattgcaata
      541 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcccata
      601 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
      661 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
      721 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
      781 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
      841 taaacgggcc ctctagatta cttgtacagc tcgtccatgc cgccggtgga gtggcggccc
      901 tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac
      961 ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc
     1021 ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc
     1081 ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc
     1141 ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg
     1201 aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag
     1261 ttcatcacgc gctcctactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg
     1321 tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc
     1381 caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg
     1441 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc
     1501 atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg
     1561 ctcaccattg ggccgggatt ttcctccacg tccccgcatg ttagaagact tcccctgccc
     1621 tcggatccca gtgtcagtct ttcaagtgga ggcagttgaa gggatcccgt gagtgagaac
     1681 tggtcctgct cggtgggctt ctccacccag gcccccagcc ccgccttgat gaaggctgtg
     1741 aacagacgcg ccttggcggc ctggtactcc tttgccgcca gcttggactc atggtacacg
     1801 ttgggcttgg taatcttgga gcgtagtaag tgggagggaa ccttgccgtg cacacgcatc
     1861 cagcgacagt acaacgcgtg cttacacagg cgggacgcgc ggcccagctc atccttccca
     1921 gtcgtggcgt tgatgacctc aatagcggag tcgcctaccg tccagttgac actgaagttg
     1981 ggggccttcc ctggctgccg tgcttctgca ttgctgatgc cactgagcaa aggcttgttg
     2041 agggtgtaga gaggtggcag gtcctctatg ttggagatcc gctggtacat ggccctggaa
     2101 aggtggtccc cgtggtaaag gctgcccagg atgatgctcg agaagtaaat gggctccacg
     2161 aaaatgctga gcagtgatcc ctggatgccc accacgttcc agcgtgcaat cttgtcactg
     2221 caggacatgg tgagcagccg ctccccttgc agcaccccgt cccacgtttg gatgctcgca
     2281 ttggagcgca ctggaatcgt cccctgacca gactctattt tggtccgtag ctgtcctctt
     2341 gctttacgat ttgggtgtct atctgctggt tcttccagga ttggctcatg tggtgagaag
     2401 attctggcat ctccacaggg agaggtgctg atgtacagat gaaactggac attctccttc
     2461 agcctaaacc cccctcgctc tgatttctga aagatggatc ttttttgatc atctttgtta
     2521 tttaagtaaa gctcaagttg tgtataaaga aatctgagca aggatctccg agatattatt
     2581 tctgcatggc agtcatttaa tgcaaggcca cgatcactca tgtattcacc attaatacat
     2641 ttgcctcctg tagaaacact tatcaccttg gcatctttaa catctgtgcc tgttgtcatg
     2701 acgactccag ccagcacttt tctgcgagcg tgaggggagg agaagttgtc ggtcaggtca
     2761 ccaaacttac ccaggaccag gcgtgagaca gcgtcagcta aaacctgcgg taaatgcagc
     2821 tgtgaccccc cgctgctgcc cccgctgctt tcaggtgttg cgctctcgct tgtgccaggt
     2881 gtctcagagc cagaggagcc tccgctagat cctccggaca cgtagatctg cagtccgccg
     2941 ttgtactgag cagaagggtg cttcacggtc agtttagaca cgtcagcggc gctgtgagcg
     3001 gctctgtact cttccaggct gtcagttttc tggtccatag ctttgtgccg gttggtggca
     3061 actttcttgc cgcccatgta gtagctccgg ttcagattgg ccttgtgaca ggcctgtcta
     3121 tcatgccttc tgaactggtg caccatgtat ggtctgccct tagggctgct caccttcttg
     3181 gcgtcggtca gtgcgctgca ggcgatgcag taggcgtcca ggtagtggtc cttagggatg
     3241 ccatgctctt cccgaaacag ataggtgtct tggccgcttg tcacgcagaa atttccgggg
     3301 aacatatcgg ccagctgatc agccaggtag gggatgatct ggttcagcac agacagggcg
     3361 tggtatttct tgttcattcc agacttcttg ctggccaggt tggcctccca ctccttatct
     3421 gtgtgcacca gtctcgcgtg ttcttcacaa aggcccactc tgttttccag tgtctcgctg
     3481 ccgttctttc ttctggggac cacgtggtgg tagtggtcga tgccgtgttt acaaaacagg
     3541 cagtgtccat cctgctgcat ggacacggcc tcttccacgc tgcctttgcc gtacagaggg
     3601 ccccgctggt actgccatct ctgaaccttg gggttgttca tggccataaa gctgaaccga
     3661 ttcagctcca gcaccacctt ggccacgggc aggatcttct gcaccttctt aaccacgttc
     3721 aggtgggtga ccagcaggtg attggcggta ggtgtcagcc atcctttagg ccgcttgcgg
     3781 ttgttgaatc tggcttcctt gttccggatg ctcttgcatg taataggctt tttgcagcca
     3841 ggcagcagtc tttgcttttc gccctcctcg aaggcggtgc cggcagcctt ggctctccgc
     3901 cgtcttttgc accgccgctt cagccttctg tgggcacgcc tatactgctt tctgtccttc
     3961 atcagcttag gcacgtcctt gtttcttgtc tcgatttggg cgttgaacac gctctctccg
     4021 ctctcggtca ccacgctcat accgatattg gttctgccag gagcaatgcc cagcaccaga
     4081 ggctgggttt cctcggcaga ctcgtaggtc agctggatgg tgaagggctt tctttccacc
     4141 actctagcct tgccttcctt gagcaagatc cgcacgtggc cgcatctggt ggtgggcatc
     4201 agtggtttgc cggacttaga gatcacgtag accacggcca tagatcccac cttccgtttc
     4261 ttttttggtc ccatggtggc agtgggttct ctagttagcc agagagctct gcttatatag
     4321 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     4381 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     4441 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     4501 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     4561 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     4621 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     4681 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     4741 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     4801 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     4861 tattaataac tagtcaataa tcaatgtcaa atgtgcgcgg aacccctatt tgtttatttt
     4921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5221 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5521 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6601 gggggcggag cctatggaaa ggcatgcaag cttgg
//
