LOCUS       Exported                5390 bp DNA     circular SYN 09-SEP-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5390)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 2023 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      208..796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(1051..1077)
                     /label=PDOTS10_RVS
                     /note="color: black; sequence: GTGGTCTCCCTATAGTGAGTCGTATTA;
                     added: 2019-07-09"
     promoter        1051..1069
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     RBS             1099..1121
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             1141..1158
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             1162..1194
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /note="color: #cc99b2"
                     /translation="MASMTGGQQMG"
     CDS             1198..1221
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /note="This feature has 2 segments:
                        1: 1198 .. 1206 / #cc99b2
                        2: 1207 .. 1221 / #cc99b2 / enterokinase site
                     Cleavage site after base 1221"
                     /translation="DLYDDDDK"
     CDS             1636..2352
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1636 .. 1638 / #00ffff
                        2: 1639 .. 1641 / #00ffff / 1a
                        3: 1642 .. 2352 / #00ffff"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2346..2385)
                     /label=pDOTS10_rvs_lin_ECFPVenus
                     /note="color: black; sequence: 
                     TGCAATACCATACTGTTCGATtccggtaccaccCTTGTAC"
     CDS             2365..3069
                     /codon_start=1
                     /label=cpAncQ binding protein
                     /note="color: #993366"
                     /translation="IEQYGIALKKGNDELLEKINKALKKLKANGTYDKIYEKWLFGKEV
                     SGGSGGSGGSGGSGDKKLRVGTDATYPPFEFKDENGELTGFDIDLAKAIAKEMGVKVEF
                     VNMDFDGLIPALQSGKIDIAISAITITDERKKTVDFSDPYYDAGLAIVVKKDNNGIKSV
                     EDLKGKRVAVQTGTTSEEYVKENFPGAGVKSFENMSEAFMELKNGRVDAVITDNPVAAD
                     YVKTSPDGKLKVV"
     primer_bind     2365..2388
                     /label=cpAncQ_ins_fwd_pETMCS3
                     /note="color: black; sequence: 
                     AGAACCTGTATTTTCAGGGTATCGAACAGTATGGTATTGCACTG"
     primer_bind     complement(3046..3069)
                     /label=cpAncQ_ins_rvs_pETMCS3
                     /note="color: black; sequence: 
                     GTACCCGGGAGCTCGAATTCTTAAACAACTTTCAGTTTACCATCCGG"
     primer_bind     3049..3090
                     /label=pDOTS10_fwd_lin_ECFPVenus
                     /note="color: black; sequence: 
                     GATGGTAAACTGAAAGTTGTTCTCgccggtaccggtggaATG"
     CDS             3088..3807
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 3088 .. 3090 / #e1ff2f
                        2: 3091 .. 3093 / #e1ff2f / 1a
                        3: 3094 .. 3807 / #e1ff2f"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     3861..3882
                     /label=pDOTS10_FWD
                     /note="color: black; sequence: CACCGCTGAGCAATAACTAGCA; 
                     added: 2019-07-09"
     terminator      3877..3924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      4021..4476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
       61 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      121 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      181 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      241 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      301 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      361 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      421 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      481 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      541 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      601 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      661 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      721 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      781 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg gcttttgctg
      841 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      901 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      961 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1021 tcattaatgc agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac
     1081 ggtttccctc tagataattt tgtttaactt taagaaggag atatacatat gcggggttct
     1141 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
     1201 ctgtacgacg atgacgataa ggatccgaaa ctgaaggtaa cagtcaacgg cactgcgtat
     1261 gacgttgacg ttgacgtcga caagtcacac gaaaacccga tgggcaccat cctgttcggc
     1321 ggcggcaccg gcggcgcgcc ggcaccggca gcaggtggcg caggcgccgg taaggccgga
     1381 gagggcgaga ttcccgctcc gctggccggc accgtctcca agatcctcgt gaaggagggt
     1441 gacacggtca aggctggtca gaccgtgctc gttctcgagg ccatgaagat ggagaccgag
     1501 atcaacgctc ccaccgacgg caaggtcgag aaggtcctgg tcaaggagcg tgacgcggtg
     1561 cagggcggtc agggtctcat caagatcggg gatctcgagc tcatcgaagg ctcgagcggt
     1621 tcggatccgg gccgcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1681 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1741 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1801 gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cagccgctac
     1861 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1921 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1981 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2041 aacatcctgg ggcacaagct ggagtacaac tacatcagcc acaacgtcta tatcaccgcc
     2101 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     2161 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2221 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2281 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2341 gagctgtaca agggtggtac cggaatcgaa cagtatggta ttgcactgaa aaaaggcaat
     2401 gatgaactgc tggaaaaaat caacaaagcc ctgaaaaaac tgaaagccaa tggcacctat
     2461 gacaaaatct atgagaaatg gctgtttggc aaagaggtga gcggtggttc tggtggttct
     2521 ggtggttctg gtggttctgg tgataaaaaa ctgcgtgttg gcaccgatgc aacctatccg
     2581 ccttttgaat ttaaagatga aaatggtgaa ctgaccggct ttgatattga tctggcaaaa
     2641 gcaattgcca aagaaatggg tgtgaaagtc gaatttgtga acatggattt tgatggtctg
     2701 attccggcac tgcagagcgg taaaattgat attgcaatta gcgccattac catcaccgat
     2761 gaacgtaaaa aaaccgtgga ttttagcgat ccgtattatg atgcaggtct ggccattgtt
     2821 gtgaaaaaag ataacaacgg catcaaaagc gtcgaagatc tgaaaggtaa acgtgttgcc
     2881 gttcagaccg gtacaaccag cgaagaatat gtgaaagaaa actttccggg tgccggtgtt
     2941 aaaagctttg aaaatatgag cgaagccttt atggaactga aaaatggtcg tgttgatgcg
     3001 gtgattaccg ataatccggt tgcagcagat tatgttaaaa ccagtccgga tggtaaactg
     3061 aaagttgttc tcgccggtac cggtggaatg gtgagcaagg gcgaggagct gttcaccggg
     3121 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3181 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     3241 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc
     3301 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3361 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3421 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3481 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3541 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     3601 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3661 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     3721 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3781 ctcggcatgg acgagctgta caagtaaaag cttgatccgg ctgctaacaa agcccgaaag
     3841 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     3901 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc tggcgtaata
     3961 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4021 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4081 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4141 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4201 gagctttacg gcacctcgac cgcaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4261 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4321 gactcttgtt ccaaactgga acaacactca accctatcgc ggtctattct tttgatttat
     4381 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaatattta
     4441 acgcgaattt taacaaaata ttaacgttta caatttcgcc tgatgcggta ttttctcctt
     4501 acgcatctgt gcggtatttc acaccgcata caggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4861 tgagcacttt taaagttctg ctatgtgata cactattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagagtg acaccacgat gcctgtagca atgccaacaa
     5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5281 actgaatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
//
