LOCUS       E5K5_pYAO_CMV_dW        5340 bp    DNA     circular SYN 02-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1..241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1567..1623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2947..2973
                     /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_E5spacer_R
     primer_bind     251..294
                     /label=xct287_E5K5_F
     misc_feature    251..266
                     /label=K5
     misc_feature    267..444
                     /label=wRNA
     primer_bind     complement(416..491)
                     /label=xct113_wRpAsig_F1
     polyA_signal    456..511
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(462..539)
                     /label=xct114_wRpAsig_F2
     primer_bind     508..527
                     /label=xct097_wRpA_GGseq
     primer_bind     complement(522..545)
                     /label=xct109_wRNAgg_F
     polyA_signal    540..764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     542..559
                     /label=xct108_TmCgg_R
     CDS             complement(802..1512)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             complement(1513..1566)
                     /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     1514..1531
                     /label=xct176_E3T2A_R
     primer_bind     complement(1548..1568)
                     /label=xct107_TmCgg_F
     primer_bind     1565..1588
                     /label=xct106_IscBgg_R
     CDS             complement(1567..1623)
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=newnucleoplasmin NLS
                     /label=nucleoplasmin NLS
     primer_bind     complement(1608..1643)
                     /label=xct168_CNLS_F
     CDS             complement(1624..2946)
                     /codon_start=1
                     /label=IscB
                     /translation="MAVVYVISKSGKPLMPTTRCGHVRILLKEGKARVVERKPFTIQLT
                     YESAEETQPLVLGIAPGRTNIGMSVVTESGESVFNAQIETRNKDVPKLMKDRKQYRRAH
                     RRLKRRCKRRRRAKAAGTAFEEGEKQRLLPGCKKPITCKSIRNKEARFNNRKRPKGWLT
                     PTANHLLVTHLNVVKKVQKILPVAKVVLELNRFSFMAMNNPKVQRWQYQRGPLYGKGSV
                     EEAVSMQQDGHCLFCKHGIDHYHHVVPRRKNGSETLENRVGLCEEHARLVHTDKEWEAN
                     LASKKSGMNKKYHALSVLNQIIPYLADQLADMFPGNFCVTSGQDTYLFREEHGIPKDHY
                     LDAYCIACSALTDAKKVSSPKGRPYMVHQFRRHDRQACHKANLNRSYYMGGKKVATNRH
                     KAMDQKTDSLEEYRAAHSAADVSKLTVKHPSAQYNGGLQIYV"
     primer_bind     1624..1643
                     /label=xct167_EIscBWT_R
     primer_bind     complement(1626..1657)
                     /label=xct177_E5dT_F
     primer_bind     1638..1669
                     /label=xct178_E5dT_R
     misc_feature    1768..1770
                     /label=L393K
     misc_feature    2140..2142
                     /label=H269A
     misc_feature    2470..2472
                     /label=V159K
     misc_feature    2536..2538
                     /label=F137K
     misc_feature    2641..2643
                     /label=M102R
     misc_feature    2767..2769
                     /label=D60A
     primer_bind     complement(2926..2966)
                     /label=xct166_EIscBWT_F
     CDS             complement(2947..2979)
                     /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     2947..2965
                     /label=xct165_NNLS_R
     misc_feature    2980..2985
                     /label=Kozak
     misc_feature    2986..3005
                     /label=arm2
     promoter        complement(3010..3213)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3039..3059)
                     /label=xct164_CMV_seq
     enhancer        3214..3593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3560..3589)
                     /label=xct103_CMVgg_F
     primer_bind     3586..3610
                     /label=xct115_YAObb_R
     promoter        3601..3705
                     /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(3706..3774,3775..4566)
                     /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             3706..3817
                     /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(3710..3731)
                     /label=xct094A_Amp_GGseq
     CDS             3818..4566
                     /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      4737..5325
                     /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     5162..5180
                     /label=ori-t-R
     primer_bind     complement(5308..5329)
                     /label=xct116_YAObb_F
     primer_bind     join(5326..5340,1..18)
                     /label=xct110_wRNAgg_R
     misc_feature    5326..5340
                     /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 tacctgtctg tggaagggct ctttcaactt gaaaaggttg aaagagcaca
      301 ggctgagaca ttcgtaaggc cgaaaggccg gacgcaccct gggatttccc cagtccccgg
      361 aactgcatag cggatgtcag ttgatgaaaa tcagataagc cagggggaac aatcacctct
      421 ctggaaacag agagagtttt ttttgttttg cagtaaataa aggaaattta ttttcattgc
      481 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatcc
      541 catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct
      601 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
      661 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      721 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg
      781 ggtttaaacg ggccctctag attacttgta cagctcgtcc atgccgccgg tggagtggcg
      841 gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc
      901 caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt
      961 ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct gcttgatctc
     1021 gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc agcccatggt
     1081 cttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta
     1141 gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc
     1201 gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca agtagtcggg
     1261 gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag gggacaggat
     1321 gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc
     1381 ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc
     1441 ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat cctcctcgcc
     1501 cttgctcacc attgggccgg gattttcctc cacgtccccg catgttagaa gacttcccct
     1561 gccctctccc tttttctttt ttgcctggcc ggcctttttc gtggccgccg gccttttgga
     1621 ccccacgtag atctgcagtc cgccgttgta ctgagcagaa gggtgcttca cggtcagttt
     1681 agacacgtca gcggcgctgt gagcggctct gtactcttcc aggctgtcag ttttctggtc
     1741 catagctttg tgccggttgg tggcaacttt cttgccgccc atgtagtagc tccggttcag
     1801 attggccttg tgacaggcct gtctatcatg ccttctgaac tggtgcacca tgtatggtct
     1861 gcccttaggg ctgctcacct tcttggcgtc ggtcagtgcg ctgcaggcga tgcagtaggc
     1921 gtccaggtag tggtccttag ggatgccatg ctcttcccga aacagatagg tgtcttggcc
     1981 gcttgtcacg cagaaatttc cggggaacat atcggccagc tgatcagcca ggtaggggat
     2041 gatctggttc agcacagaca gggcgtggta tttcttgttc attccagact tcttgctggc
     2101 caggttggcc tcccactcct tatctgtgtg caccagtctc gcgtgttctt cacaaaggcc
     2161 cactctgttt tccagtgtct cgctgccgtt ctttcttctg gggaccacgt ggtggtagtg
     2221 gtcgatgccg tgtttacaaa acaggcagtg tccatcctgc tgcatggaca cggcctcttc
     2281 cacgctgcct ttgccgtaca gagggccccg ctggtactgc catctctgaa ccttggggtt
     2341 gttcatggcc ataaagctga accgattcag ctccagcacc accttggcca cgggcaggat
     2401 cttctgcacc ttcttaacca cgttcaggtg ggtgaccagc aggtgattgg cggtaggtgt
     2461 cagccatcct ttaggccgct tgcggttgtt gaatctggct tccttgttcc ggatgctctt
     2521 gcatgtaata ggctttttgc agccaggcag cagtctttgc ttttcgccct cctcgaaggc
     2581 ggtgccggca gccttggctc tccgccgtct tttgcaccgc cgcttcagcc ttctgtgggc
     2641 acgcctatac tgctttctgt ccttcatcag cttaggcacg tccttgtttc ttgtctcgat
     2701 ttgggcgttg aacacgctct ctccgctctc ggtcaccacg ctcataccga tattggttct
     2761 gccaggagca atgcccagca ccagaggctg ggtttcctcg gcagactcgt aggtcagctg
     2821 gatggtgaag ggctttcttt ccaccactct agccttgcct tccttgagca agatccgcac
     2881 gtggccgcat ctggtggtgg gcatcagtgg tttgccggac ttagagatca cgtagaccac
     2941 ggccatagat cccaccttcc gtttcttttt tggtcccatg gtggcagtgg gttctctagt
     3001 tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     3061 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     3121 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     3181 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     3241 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     3301 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     3361 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     3421 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     3481 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     3541 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaaatgtg
     3601 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3661 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3721 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3781 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3841 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3901 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3961 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4021 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4081 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4141 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4201 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4261 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4321 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4381 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4441 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4501 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4561 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4621 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4681 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4741 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4801 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4861 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4921 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4981 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5041 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5101 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5161 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5221 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5281 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaggcat gcaagcttgg
//
