LOCUS       E5TS3_30_pYAO_CM        5829 bp    DNA     circular SYN 08-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5829)
  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..5829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2368..2394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3718..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          5589..5829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=459_Sa_HLE7_F
     primer_bind     1..20
                     /label=469_SpHLE7_F
     misc_feature    1..15
                     /label=MCS
     primer_bind     12..33
                     /label=xct116_YAObb_F
     rep_origin      16..604
                     /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     complement(161..179)
                     /label=ori-t-R
     primer_bind     complement(523..542)
                     /label=xct099_ori_GGseq
     primer_bind     complement(600..619)
                     /label=403_Spdsg_insn3_R
     CDS             complement(join(775..1566,1567..1635))
                     /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             complement(775..1523)
                     /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"
     primer_bind     775..797
                     /label=402_Spdsg_insn3_F
     CDS             complement(1524..1635)
                     /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     1610..1631
                     /label=xct094A_Amp_GGseq
     promoter        complement(1636..1740)
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     primer_bind     complement(1731..1755)
                     /label=xct115_YAObb_R
     enhancer        1748..2127
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1752..1781
                     /label=xct103_CMVgg_F
     promoter        2128..2331
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2282..2302
                     /label=xct164_CMV_seq
     misc_feature    2336..2355
                     /label=arm2
     primer_bind     complement(2342..2361)
                     /label=477_rmIscB_R
     misc_feature    2356..2361
                     /label=Kozak
     CDS             2362..2394
                     /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     2368..2389
                     /label=461_HLSaTT_F2
     primer_bind     2375..2415
                     /label=xct166_EIscBWT_F
     primer_bind     complement(2376..2394)
                     /label=xct165_NNLS_R
     CDS             2395..3717
                     /codon_start=1
                     /label=IscB
                     /translation="MAVVYVISKSGKPLMPTTRCGHVRILLKEGKARVVERKPFTIQLT
                     YESAEETQPLVLGIAPGRTNIGMSVVTESGESVFNAQIETRNKDVPKLMKDRKQYRRAH
                     RRLKRRCKRRRRAKAAGTAFEEGEKQRLLPGCKKPITCKSIRNKEARFNNRKRPKGWLT
                     PTANHLLVTHLNVVKKVQKILPVAKVVLELNRFSFMAMNNPKVQRWQYQRGPLYGKGSV
                     EEAVSMQQDGHCLFCKHGIDHYHHVVPRRKNGSETLENRVGLCEEHARLVHTDKEWEAN
                     LASKKSGMNKKYHALSVLNQIIPYLADQLADMFPGNFCVTSGQDTYLFREEHGIPKDHY
                     LDAYCIACSALTDAKKVSSPKGRPYMVHQFRRHDRQACHKANLNRSYYMGGKKVATNRH
                     KAMDQKTDSLEEYRAAHSAADVSKLTVKHPSAQYNGGLQIYV"
     primer_bind     2398..2421
                     /label=xct197_F
     primer_bind     complement(2554..2572)
                     /label=xct313_ER1ruv_R
     misc_feature    2572..2574
                     /label=D60A
     misc_feature    2698..2700
                     /label=M102R
     misc_feature    2803..2805
                     /label=F137K
     misc_feature    2869..2871
                     /label=V159K
     misc_feature    3199..3201
                     /label=H269A
     primer_bind     3459..3477
                     /label=xct175_EIscB_seq
     misc_feature    3571..3573
                     /label=L393K
     primer_bind     complement(3670..3713)
                     /label=xct170_EdTop_R
     primer_bind     complement(3672..3703)
                     /label=xct178_E5dT_R
     primer_bind     3672..3691
                     /label=xct096_IscBC_GGseq
     primer_bind     3684..3715
                     /label=xct177_E5dT_F
     primer_bind     3694..3715
                     /label=xct169_EdTop_F
     primer_bind     3694..3714
                     /label=xct174_ES1lin_F
     primer_bind     complement(3698..3774)
                     /label=xct112_IscBNLSC_R
     primer_bind     3698..3733
                     /label=xct168_CNLS_F
     primer_bind     complement(3698..3717)
                     /label=xct167_EIscBWT_R
     primer_bind     complement(3699..3717)
                     /label=479_rmADAR_R
     CDS             3718..3774
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=newnucleoplasmin NLS
                     /label=nucleoplasmin NLS
     CDS             3775..3828
                     /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     3809..3846
                     /label=553_GFPin_F
     primer_bind     complement(3810..3827)
                     /label=xct176_E3T2A_R
     primer_bind     3826..3846
                     /label=560_ts5end_R
     CDS             3829..4113
                     /codon_start=1
                     /label=GFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLT*GVQCFSRYPDHMKQHDFFKSAMPEGYVQ"
     primer_bind     3829..3844
                     /label=xct065_GexGFP_F
     regulatory      3829..3832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3832..3849
                     /label=375_EGFP_F
     primer_bind     4002..4019
                     /label=408_GFPW58X_F
     primer_bind     complement(4009..4028)
                     /label=557_Ystop_R
     primer_bind     4029..4046
                     /label=556_Ystop_F
     misc_feature    4114..4850
                     /label=LMNA Intron
     misc_feature    complement(4331..4380)
                     /label=3' best guide
     misc_feature    complement(4575..4604)
                     /label=5' best guide
     primer_bind     4575..4602
                     /label=559_ts5end_F
     CDS             4851..5285
                     /codon_start=1
                     /label=GFP
                     /translation="ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH
                     KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDN
                     HYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(5262..5282)
                     /label=376_EGFP_R
     primer_bind     complement(5264..5285)
                     /label=555_GFPin_Ra
     primer_bind     5266..5307
                     /label=554_intronbb_F
     primer_bind     complement(5266..5285)
                     /label=555_GFPin_R
     primer_bind     5286..5306
                     /label=394_PuroC2_F
     primer_bind     5298..5317
                     /label=354_EHLSp9bb_F
     polyA_signal    5323..5547
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5323..5342
                     /label=535_Spsgdual_F
     primer_bind     5548..5579
                     /label=xct114_wRpAsig_F2
     primer_bind     complement(5560..5579)
                     /label=xct097_wRpA_GGseq
     polyA_signal    5576..5579
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5580..5599
                     /label=xct228_E5spacer_R
     primer_bind     complement(5580..5599)
                     /label=560_ts5end_R
     promoter        complement(5589..5829)
                     /label=U6 promoter RNA polymerase III promoter for hum
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(5808..5829)
                     /label=382_SpsgGG_R
     primer_bind     complement(5809..5829)
                     /label=hU6-F
ORIGIN
        1 ccaagcttgc atgcctttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
       61 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1741 cacatttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1801 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1861 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1921 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1981 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2041 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2101 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2161 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2221 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2281 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     2341 actagagaac ccactgccac catgggacca aaaaagaaac ggaaggtggg atctatggcc
     2401 gtggtctacg tgatctctaa gtccggcaaa ccactgatgc ccaccaccag atgcggccac
     2461 gtgcggatct tgctcaagga aggcaaggct agagtggtgg aaagaaagcc cttcaccatc
     2521 cagctgacct acgagtctgc cgaggaaacc cagcctctgg tgctgggcat tgctcctggc
     2581 agaaccaata tcggtatgag cgtggtgacc gagagcggag agagcgtgtt caacgcccaa
     2641 atcgagacaa gaaacaagga cgtgcctaag ctgatgaagg acagaaagca gtataggcgt
     2701 gcccacagaa ggctgaagcg gcggtgcaaa agacggcgga gagccaaggc tgccggcacc
     2761 gccttcgagg agggcgaaaa gcaaagactg ctgcctggct gcaaaaagcc tattacatgc
     2821 aagagcatcc ggaacaagga agccagattc aacaaccgca agcggcctaa aggatggctg
     2881 acacctaccg ccaatcacct gctggtcacc cacctgaacg tggttaagaa ggtgcagaag
     2941 atcctgcccg tggccaaggt ggtgctggag ctgaatcggt tcagctttat ggccatgaac
     3001 aaccccaagg ttcagagatg gcagtaccag cggggccctc tgtacggcaa aggcagcgtg
     3061 gaagaggccg tgtccatgca gcaggatgga cactgcctgt tttgtaaaca cggcatcgac
     3121 cactaccacc acgtggtccc cagaagaaag aacggcagcg agacactgga aaacagagtg
     3181 ggcctttgtg aagaacacgc gagactggtg cacacagata aggagtggga ggccaacctg
     3241 gccagcaaga agtctggaat gaacaagaaa taccacgccc tgtctgtgct gaaccagatc
     3301 atcccctacc tggctgatca gctggccgat atgttccccg gaaatttctg cgtgacaagc
     3361 ggccaagaca cctatctgtt tcgggaagag catggcatcc ctaaggacca ctacctggac
     3421 gcctactgca tcgcctgcag cgcactgacc gacgccaaga aggtgagcag ccctaagggc
     3481 agaccataca tggtgcacca gttcagaagg catgatagac aggcctgtca caaggccaat
     3541 ctgaaccgga gctactacat gggcggcaag aaagttgcca ccaaccggca caaagctatg
     3601 gaccagaaaa ctgacagcct ggaagagtac agagccgctc acagcgccgc tgacgtgtct
     3661 aaactgaccg tgaagcaccc ttctgctcag tacaacggcg gactgcagat ctacgtgggg
     3721 tccaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gggagagggc
     3781 aggggaagtc ttctaacatg cggggacgtg gaggaaaatc ccggcccaat ggtgagcaag
     3841 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3901 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3961 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4021 ctgacctaag gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     4081 ttcaagtccg ccatgcccga aggctacgtc caggtagccg ccgctgaggc cgagcctgca
     4141 ctggggccac ccagccaggc ctgggggcag cctctcccca gcctccccgt gccaaaaatc
     4201 ttttcattaa agaatgtttt ggaactttac tcgctggcct ggcctttctt ctctctcctc
     4261 cctatacctt gaacagggaa cccaggtgtc tgggtgccct actctggtaa ggaagggagt
     4321 gggaactttc tgatgccatg gaatattcct gtgggagcag tggacaaggg tctggatttg
     4381 tcttctggga aagggagggg aggacagacg tggggcatgc ccgccctgcc tctctccccc
     4441 attcttgttg catgcatatc ctctcatttc cctcattttt cctgcaagaa tgttctctct
     4501 cattcctgac cgcccctcca ctccaattaa tagtgcatgc ctgctgccct acaagcttgc
     4561 tcccgttctc tcttcttttc ctcttaagct cagagtagct agaacagagt cagagtcact
     4621 gctctggttc tctgtcccca agtcttcctg agccttctcc ccttttatgt cttccctctc
     4681 ctcctccggg cccctagcct cccaaacccc cattgcccgc tggctccttg ggcacagaac
     4741 cacaccttcc tgcctggcgg ctgggagcct gcaggagcct ggagcctggt tgggcctgag
     4801 tggtcagtcc cagactcgcc gtcccgcctg agccttgtct cccttcccag gagcgcacca
     4861 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     4921 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     4981 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5041 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5101 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5161 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5221 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5281 agtaatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     5341 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5401 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5461 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5521 atgctgggga tgcggtgggc tctatgggat ttgccctccc atatgtcctt ccgagtgagc
     5581 ggtgtttcgt cctttccaca agatatataa agccaagaaa tcgaaatact ttcaagttac
     5641 ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact acccaagaaa
     5701 ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag tcaaattaat
     5761 tccaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa
     5821 taggccctc
//
