LOCUS       Exported                4675 bp DNA     circular SYN 19-AUG-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4675)
  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..4675
                     /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..804
                     /label=InFu_CENP-O_F
                     /note="color: black; sequence: 
                     attccccggtgctagATGGAGTCTGCAAATACTTTGTGTCC"
     primer_bind     764..790
                     /label=InFu_Rn-O-80_F
                     /note="color: black; sequence: attccccggtgctagATGGAGTCTGCA"
     misc_feature    779..1030
                     /label=MmCENP-O
                     /note="color: #33cccc"
     primer_bind     complement(997..1033)
                     /label=InFu_Rn-O-80_R
                     /note="color: black; sequence: 
                     CGTGGTTTTGCTTGAGTCTTCTTTGGCAGGAGGTTCT"
     misc_feature    1019..1675
                     /label=RnCenp-O
                     /note="color: #339966"
     primer_bind     1019..1042
                     /label=O-80_Vector_F
                     /note="color: black; sequence: TCAAGCAAAACCACGCTTAGTGAA"
     primer_bind     1172..1195
                     /label=RnCENP-O_seq1
                     /note="color: black; sequence: CTACTAGATTCCTACTTTGTGGAC"
     primer_bind     complement(1654..1690)
                     /label=InFu_RnCENP-O_R
                     /note="color: black; sequence: 
                     tccgccaccgACCGGGGAGACCAGGTTCAAATGCAGC"
     primer_bind     complement(1654..1675)
                     /label=RnCENP-O_R
                     /note="color: black; sequence: GGAGACCAGGTTCAAATGCAGC"
     primer_bind     1676..1689
                     /label=HOLI_Vector_F
                     /note="color: black; sequence: CCGGTcggtggcgg"
     misc_feature    1682..1696
                     /label=5Gly
                     /note="color: #ffff00"
     primer_bind     1682..1696
                     /label=5Gly_seq_1
                     /note="color: black; sequence: ggtggcggaggtggc"
     CDS             1697..2410
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 2 segments:
                        1: 1697 .. 1699 / #05fd14 / 1a
                        2: 1700 .. 2410 / #05fd14"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2391..2429)
                     /label=Fragment.REV2
                     /note="color: black; sequence: 
                     tggtttagtggtaacctcacttgtacagctcgtccatgc"
     promoter        complement(2655..2673)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2691..2707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2715..2731
                     /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(2739..2769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2739 .. 2745 / #ffffff / -10
                        2: 2746 .. 2763 / #ffffff
                        3: 2764 .. 2769 / #ffffff / -35"
     protein_bind    2784..2805
                     /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(3093..3682)
                     /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 ggagtctgca aatactttgt gtccaggcag gaagtgcaaa ggaggtgttt tagctcacct
      841 ggaaagacta gaggctcaaa cgaacatatc gaatagaaag tctgaagaac cagcagttcg
      901 taaaaaggaa agttctctta gaaccaagat tcgagaactg cggcaacaac gagataagct
      961 gagggcagaa gtgaagcagt ggggagccag agttaaagaa cctcctgcca 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 agagaagcgc cctgtgcaac ttgctgtcat tcacctataa
     1441 agtggaacac gggacccagg ccttctcctt tagtgccagg ttgctgtatg aggacctcac
     1501 agcagctctt cccacaggag tcactgtgac aaggccagca ggtgtggaag catcgtcccc
     1561 tcattgggaa gagcacagag catcccatga aatgctcttc cgtaccaaga ccctgcacca
     1621 ggcgtttgct tcattttcca aagagacaga aatgctgcat ttgaacctgg tctccccggt
     1681 cggtggcgga ggtggcgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1741 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1801 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1861 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1921 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1981 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2041 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2101 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2161 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2221 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2281 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2341 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2401 gctgtacaag tgaggttacc actaaaccag cctcaagaac acccgaatgg agtctctaag
     2461 ctacataata ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat
     2521 ctgctcctaa taaaaagaaa gtttcttcac attctaaaaa aaaaaaaaaa aaaaaaaaaa
     2581 aaaaaaaaac cccccccccc cccccctgca ggcgcgccgc cggcgattta aattaattaa
     2641 tctagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg
     2701 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2761 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2821 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2881 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2941 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3001 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3421 cacgacctta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4021 atcagcaata aaccagccag cccggaaggg ccgagcgcag aagtggtcct gcaaacttta
     4081 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4141 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4201 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4261 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4321 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4381 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4441 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4501 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4561 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4621 tttactttca ccagcgtttc tgggtgagcc ctttcaccag cgtttctggg tgagc
//
