LOCUS       Exported                4672 bp DNA     circular SYN 19-AUG-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4672)
  AUTHORS   Covid-19 WFH
  TITLE     Direct Submission
  JOURNAL   Exported Jul 3, 2023 from SnapGene 6.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4672
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        complement(63..167)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     551..570
                     /label=QCH_seq_primer_1
                     /note="color: black; sequence: CCTCTTCGCTATTACGCCAG"
     primer_bind     641..657
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        664..682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     5'UTR           714..756
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     complement(756..778)
                     /label=HOLI_Vector_R
                     /note="color: black; sequence: ctagcaccggggaattgatctgc"
     primer_bind     764..806
                     /label=InFu_RnCENP-O_F
                     /note="color: black; sequence: 
                     attccccggtgctagATGGAATCTGCAAATTCTTTGTGTCCAG"
     misc_feature    779..1387
                     /label=RnCenp-O
                     /note="color: #339966"
     primer_bind     779..805
                     /label=RnCENP-O_F
                     /note="color: black; sequence: ATGGAATCTGCAAATTCTTTGTGTCCA"
     primer_bind     1172..1195
                     /label=RnCENP-O_seq1
                     /note="color: black; sequence: CTACTAGATTCCTACTTTGTGGAC"
     primer_bind     complement(1365..1387)
                     /label=O-298-Vector_R
                     /note="color: black; sequence: GAGGAAAGCGCAGAAGTCACTCT"
     misc_feature    1373..1672
                     /label=MmCENP-O
                     /note="color: #33cccc"
     primer_bind     1373..1402
                     /label=InFu_Rn-O-298_F
                     /note="color: black; sequence: 
                     TTCTGCGCTTTCCTCACGGGACCTTTGCAG"
     primer_bind     complement(1651..1687)
                     /label=InFu_CENP-O_R
                     /note="color: black; sequence: 
                     tccgccaccgACCGGGGAGACCAGGTTCAAATGCAGC"
     primer_bind     complement(1661..1687)
                     /label=InFu_Rn-O-298_R
                     /note="color: black; sequence: tccgccaccgACCGGGGAGACCAGGTT"
     primer_bind     1673..1686
                     /label=HOLI_Vector_F
                     /note="color: black; sequence: CCGGTcggtggcgg"
     misc_feature    1679..1693
                     /label=5Gly
                     /note="color: #ffff00"
     primer_bind     1679..1693
                     /label=5Gly_seq_1
                     /note="color: black; sequence: ggtggcggaggtggc"
     CDS             1694..2407
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 2 segments:
                        1: 1694 .. 1696 / #05fd14 / 1a
                        2: 1697 .. 2407 / #05fd14"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2388..2426)
                     /label=Fragment.REV2
                     /note="color: black; sequence: 
                     tggtttagtggtaacctcacttgtacagctcgtccatgc"
     promoter        complement(2652..2670)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2688..2704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2712..2728
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2736..2766)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2736 .. 2742 / #ffffff / -10
                        2: 2743 .. 2760 / #ffffff
                        3: 2761 .. 2766 / #ffffff / -35"
     protein_bind    2781..2802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(3090..3679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 aaacaggaag gcaaatgccg aaaaaaggga ataagggcga cacggaaatg ttgaatactc
       61 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
      121 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
      181 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
      241 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
      301 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
      361 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
      421 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
      481 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      541 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      601 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
      661 ttgtaatacg actcactata gggcgaatta attcgagctc ggtacccagc ttgcttgttc
      721 tttttgcaga agctcagaat aaacgctcaa ctttggcaga tcaattcccc ggtgctagat
      781 ggaatctgca aattctttgt gtccaggcac agagtgtaaa ggaggtgttt tagctcacct
      841 ggaaagacta gaggctcaaa tgaacagatc gaatagaaag tctgaagaac caacagttcg
      901 tgaaaaggaa agttctctta gaaccaagat tcgagaactg cagcaacagc gagataggct
      961 gagggtggaa gtgaagcagt ggcgagccag agttaaagaa tcttctgcca aagaagactc
     1021 aagcaaaacc acgcttagtg aacaggaagt tctggaaaag cagtggagaa acctgaaagc
     1081 cattctgcag gcatatcgat ttacagggct cagcgggaag ttgaccaacc gtagagtctg
     1141 tgtgtacatc agcaccgcct ttgaggggaa cctactagat tcctactttg tggacctcgc
     1201 catagagaaa ccactccgga ttcatcacca ttccatccca gccttcatcc ctctggaaaa
     1261 gatagccaca gcatacctac agactgacgt tcagcgcctc ctgtttagtc tctcggagta
     1321 cctaaatgcc tacgtgggaa ggaagtatca ggcagaccaa cttgagagtg acttctgcgc
     1381 tttcctcacg ggacctttgc agagaaacgc cctgtgcaac ttgctgtcgt tcacctataa
     1441 agtggaacag agatgccaga ccttctcctt cagtgccagg ttgctgtatg aggaccccac
     1501 agcagctctt cccacgaatg tcactgtgac aaggccaggt gtggaggcat catcccctcc
     1561 ttgggaagag cacagagcat cccatcagat gctcttccgt accaagcccc tgcacaaggt
     1621 gtttgcttca ttttccaaag aaaccgaaaa gctgcatttg aacctggtct ccccggtcgg
     1681 tggcggaggt ggcgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1741 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1801 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1861 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1921 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1981 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2041 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2101 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2161 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2221 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2281 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2341 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2401 gtacaagtga ggttaccact aaaccagcct caagaacacc cgaatggagt ctctaagcta
     2461 cataatacca acttacactt tacaaaatgt tgtcccccaa aatgtagcca ttcgtatctg
     2521 ctcctaataa aaagaaagtt tcttcacatt ctaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2581 aaaaaacccc cccccccccc ccctgcaggc gcgccgccgg cgatttaaat taattaatct
     2641 agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt
     2701 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2761 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2821 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2881 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2941 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3421 gaccttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4021 agcaataaac cagccagccc ggaagggccg agcgcagaag tggtcctgca aactttatcc
     4081 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4141 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4201 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4261 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4321 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4381 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4441 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4501 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4561 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4621 actttcacca gcgtttctgg gtgagccctt tcaccagcgt ttctgggtga gc
//
