LOCUS       Exported                5406 bp ds-DNA     circular SYN 05-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  AUTHORS   MPIBPC
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 11, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5406
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     272..288
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        295..313
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           345..387
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    411..435
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    411..418
                     /label=recomb left left part
     misc_feature    419..428
                     /label=Recomb here for pDest vector
     CDS             456..1907
                     /codon_start=1
                     /label=bNup98
                     /translation="MFNKSFGTPFGGGTGGFGTTSTFGQNTGFGTTSGGAFGTSAFGSS
                     NNTGGLFGNSQTKPGGLFGTNSFSQPATSTSTGFGFGTSTGTSNSLFGTASTGTSLFSS
                     QNNAFAQNKPTGFGNFGTSTSSGGLFGTTNTTSNPFGSTSGSLFGPSSFTAAPTGTTIK
                     FNPPTGTDTMVKAGVSTNISTKHQCITAMKEYESKSLEELRLEDYQANRKGPQNQVGAG
                     TTTGLFGSSPATSSATGLFSSSTTNSGFAYGQNKTAFGTSTTGFGTNPGGLFGQQNQTT
                     SLFSKPFAQPTTTQTTGFSFGNTSTLGQPSTNTMGLFGVTQASQPGGLFGTATNTSTGT
                     AFGTGASLFGQTSTGFGAVGSTLFGNNKLTTFGTSTTSAPSFGTTSGGLFGFGTNTSGN
                     SIFGSKPGPGTLGTGLGAGFGTALGAGQASLFGNNQPKLGGPLGTGAFGAPGFNTTTAT
                     LGFGAPQAPVALTDPNASAAQQAVLQ"
     protein_bind    complement(1924..1948)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1924..1935
                     /label=recomb until here for Pdest vector
     misc_feature    1936..1947
                     /label=recom right right def
     CDS             1995..2015
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      2025..2034
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2031..2747
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             2748..2768
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     3'UTR           2813..2938
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(3043..3061)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3079..3095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3103..3119
                     /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(3127..3157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3172..3193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3481..4070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4541..4919)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     promoter        complement(5100..5204)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
       61 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      121 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      181 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      241 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
      301 gactcactat agggcgaatt aattcgagct cggtacccag cttgcttgtt ctttttgcag
      361 aagctcagaa taaacgctca actttggcag atcaattccc cggtgctagc acaagtttgt
      421 acaaaaaagc aggctccgcg gccgccccct tcaccatgtt taataagtca tttggaacgc
      481 cctttggggg tggcactggt ggctttggca caacttcaac atttgggcag aatactggat
      541 ttggcactac tagtggaggg gcatttggaa catctgcatt tggttctagc aacaatactg
      601 gaggcctgtt tggaaattca cagaccaaac caggaggctt atttggtacc aattcattta
      661 gccagccagc tacctccaca agcactggct ttgggtttgg cacatcaaca ggaacatcaa
      721 atagcttgtt tggaactgca agcacaggaa ccagcctctt ctcatcccaa aacaatgcct
      781 ttgcacaaaa taagccaacg ggctttggga attttggaac aagtaccagc agtgggggac
      841 tttttggaac tacaaacacc acctctaatc cttttggtag cacatctggc tctctctttg
      901 ggccaagtag ttttacagct gctcctactg ggactactat taagtttaac cctccaactg
      961 gtactgatac tatggttaaa gctggagtta gcactaacat cagtaccaag caccagtgta
     1021 ttactgctat gaaagaatat gaaagcaagt cactagagga gcttcgttta gaggattatc
     1081 aggccaaccg gaaaggccca cagaatcagg tgggagcagg taccacgact ggcttgtttg
     1141 ggtcttctcc agccacctcc agtgcaacag gactcttcag ctcttccaca acgaattcag
     1201 gctttgcata tggtcagaac aaaacagcct ttggaactag tacaactgga tttggaacaa
     1261 atcctggtgg tctctttggc caacagaatc aaactaccag tctcttcagc aaaccatttg
     1321 cccagcccac aaccacccag accactggtt tttcctttgg taataccagc acactaggac
     1381 agccaagcac caacaccatg ggtctatttg gagtaactca agcctcacag ccaggaggtc
     1441 tttttgggac agctacaaac accagcactg ggacagcatt tggaacagga gctagtctct
     1501 ttgggcagac cagtactgga tttggtgctg ttggttcgac cctgtttgga aataacaagc
     1561 ttactacgtt tggaaccagc acaaccagtg ctccttcatt tggtacaacc agtggcgggc
     1621 tctttggatt cggcacaaat accagtggga atagtatttt tggaagtaaa ccaggacctg
     1681 gcactctggg gactgggctt ggcgcaggat ttggaacagc tcttggtgct ggacaggcat
     1741 ctttgtttgg gaacaaccaa cctaagctcg gagggcccct tggtacagga gcctttgggg
     1801 cccctggatt taatacaacg acagccactt tgggctttgg agccccccag gccccagtag
     1861 ctttgacaga tccaaacgct tccgctgccc agcaggctgt tctccaaaag ggtgggcgcg
     1921 ccgacccagc tttcttgtac aaagtggtat ctcgagctca agcttcgaat tctgcagtcg
     1981 acggcccggg atccccaaaa aagaagagaa aggtagaaga ccccgccacc atggtgagca
     2041 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2101 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     2161 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2221 ccttcggcta cggcgtggcc tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2281 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg
     2341 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2401 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2461 acaacttcaa cagccactac gtctatatca cggccgacaa gcagaagaac tgcatcaagg
     2521 ctaacttcaa gatccgccac aacgttgagg acggcagcgt gcagctcgcc gaccactacc
     2581 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc
     2641 atcagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2701 tcgtgaccgc cgccgggatt acacatggca tggacgagct gtacaagcca aaaaagaaga
     2761 gaaaggtaga atctagatct agataaatga tagcttgatc tggttaccac taaaccagcc
     2821 tcaagaacac ccgaatggag tctctaagct acataatacc aacttacact ttacaaaatg
     2881 ttgtccccca aaatgtagcc attcgtatct gctcctaata aaaagaaagt ttcttcacat
     2941 tctaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaccc cccccccccc cccctgcagg
     3001 cgcgccgccg gcgatttaaa ttaattaatc tagcttgagt attctatagt gtcacctaaa
     3061 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3121 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3181 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3241 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3781 ctatcgtctt gagtccaacc cggtaagaca cgaccttatc gccactggca gcagccactg
     3841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc cggaagggcc
     4441 gagcgcagaa gtggtcctgc aaactttatc cgcctccatc cagtctatta attgttgccg
     4501 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4561 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4621 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4681 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4741 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4801 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4861 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4921 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4981 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5041 caggaaggca aatgccgaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5221 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5281 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5341 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5401 cagggc
//
