LOCUS       pGEMHE-Trim21-OL        4684 bp DNA     circular SYN 28-FEB-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4684)
  AUTHORS   Covid-19 WFH
  TITLE     Direct Submission
  JOURNAL   Exported Mar 24, 2025 from SnapGene 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4684
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        complement(63..167)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(386..396)
                     /label=InFu_MmCENP-P_F
                     /note="color: black; sequence: 
                     attccccggtgctagATGGATTCCGAGACCCGC"
     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(757..781)
                     /label=Vector_Trim21_R
                     /note="color: black; sequence: CATctagcaccggggaattgatctg"
     primer_bind     764..781
                     /label=InFu_MmCENP-P_F
                     /note="color: black; sequence: 
                     attccccggtgctagATGGATTCCGAGACCCGC"
     primer_bind     767..801
                     /label=InFu_Trim21_F
                     /note="color: black; sequence: 
                     ccccggtgctagATGtccggactcagatctcgagc"
     CDS             818..2203
                     /codon_start=1
                     /label=MmTrim21 (missing first 8 aa)
                     /note="color: #c0c0c0"
                     /translation="MSLEKMWEEVTCSICLDPMVEPMSIECGHCFCKECIFEVGKNGGS
                     SCPECRQQFLLRNLRPNRHIANMVENLKQIAQNTKKSTQETHCMKHGEKLHLFCEEDGQ
                     ALCWVCAQSGKHRDHTRVPIEEAAKVYQEKIHVALEKLRKGKELAEKMEMDLTMQRTDW
                     KRNIDTQKSRIHAEFALQNSLLAQEEQRQLQRLEKDQREYLRLLGKKEAELAEKNQALQ
                     ELISELERRIRGSELELLQEVRIILERSGSWNLDTLDIDAPDLTSTCPVPGRKKMLRTC
                     WVHITLDRNTANSWLIISKDRRQVRMGDTHQNVSDNKERFSNYPMVLGAQRFSSGKMYW
                     EVDVTQKEAWDLGVCRDSVQRKGQFSLSPENGFWTIWLWQDSYEAGTSPQTTLHIQVPP
                     CQIGIFVDYEAGVVSFYNITDHGSLIYTFSECVFAGPLRPFFNVGFNYSGGNAAPLKLC
                     PLKM"
     primer_bind     complement(2176..2218)
                     /label=InFu_Trim21_R
                     /note="color: black; sequence: 
                     TCCGCCACCgACCGGcatctttagtggacagagctttagaggc"
     primer_bind     2204..2217
                     /label=Vector_Trim21_F
                     /note="color: black; sequence: CCGGTcGGTGGCGG"
     misc_feature    2210..2224
                     /label=5Gly
                     /note="color: #ffff00"
     CDS             2225..2266
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /note="color: #cc99b2"
                     /translation="SGFANELGPRLMGK"
     primer_bind     2360..2370
                     /label=Vector_Trim21_F
                     /note="color: black; sequence: CCGGTcGGTGGCGG"
     misc_feature    2363..2377
                     /label=5Gly
                     /note="color: #ffff00"
     CDS             2378..2419
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /note="color: #cc99b2"
                     /translation="SGFANELGPRLMGK"
     promoter        complement(2664..2682)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2700..2716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2724..2740
                     /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(join(2748..2754,2755..2772,2773..2778))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2748 .. 2754 / #ffffff / -10
                        2: 2755 .. 2772 / #ffffff
                        3: 2773 .. 2778 / #ffffff / -35"
     protein_bind    2793..2814
                     /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(3102..3691)
                     /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 gtccggactc agatctcgag ctcaagcttc gaattcaatg tctctggaaa agatgtggga
      841 ggaggtcacc tgttctatct gcctggatcc catggtggag cctatgagta tcgaatgtgg
      901 ccattgcttt tgcaaggaat gcatttttga agttgggaag aatgggggca gttcatgtcc
      961 cgagtgccgg caacagtttc tgctccgaaa cctcaggccc aatagacata tagccaacat
     1021 ggtggaaaac cttaaacaga tagcccagaa taccaagaag agtacccagg aaacgcactg
     1081 catgaagcat ggagagaagc ttcacctatt ctgtgaggaa gatgggcagg ccctttgctg
     1141 ggtgtgtgcc cagtctggga aacaccggga ccacaccagg gtccctattg aagaggctgc
     1201 taaggtatac caggagaaga tccacgtggc tttagaaaaa ctgagaaagg ggaaagagtt
     1261 ggccgagaag atggaaatgg atctcacgat gcaaagaaca gactggaaga ggaacattga
     1321 cacccagaag tcgaggattc acgcagagtt cgcacttcag aatagcttgc tggctcagga
     1381 ggagcagagg cagctgcaga ggctggagaa ggatcaaagg gagtacctga gactcctggg
     1441 gaagaaggag gctgagctgg ctgagaagaa ccaggccctg caggagctga tctcagagct
     1501 ggagaggagg attcgtggtt cagagctgga gctactgcag gaggtgagga tcatcctgga
     1561 aaggagtgga tcctggaacc tggacacgtt agatattgac gccccagacc taacaagcac
     1621 atgccctgtg ccagggcgga agaagatgct gaggacgtgt tgggttcata ttactctgga
     1681 tcgcaacacc gccaactcat ggctcatcat ctcaaaggat cggagacaag tgaggatggg
     1741 agacacccat cagaacgtgt ctgacaataa ggagaggttt agtaattacc ccatggtgct
     1801 aggtgcccag agattctcct ctgggaagat gtactgggag gtagatgtga ctcaaaagga
     1861 ggcctgggat ctgggggttt gcagagattc tgttcagagg aaagggcagt tttcactcag
     1921 tcccgagaat ggcttctgga ccatttggtt atggcaagac agctatgagg ctggtaccag
     1981 tcctcagacc accctccaca ttcaagtacc tccatgccaa attgggatct ttgtggacta
     2041 tgaggctggc gttgtctcct tctacaacat aactgaccat ggctccctca tttacacctt
     2101 ctcggagtgt gtttttgctg gacctctgcg acctttcttc aatgttggtt tcaattatag
     2161 tgggggaaat gcagcgcctc taaagctctg tccactaaag atgccggtcg gtggcggagg
     2221 tggcagcgga ttcgctaatg agctgggacc gaggcttatg ggaaagtgag gttaccacta
     2281 aaccagcctc aagaacaccc gaatgggctg gttaccacta aaccagcctc aagaacagaa
     2341 ccgtcaagat gagaagaccg tcggtggcgg aggtggcagc ggattcgcta atgagctggg
     2401 accgaggctt atgggaaagt gaggttacca ctaaaccagc ctcaagaaca cccgaatgga
     2461 gtctctaagc tacataatac caacttacac tttacaaaat gttgtccccc aaaatgtagc
     2521 cattcgtatc tgctcctaat aaaaagaaag tttcttcaca ttctaaaaaa aaaaaaaaaa
     2581 aaaaaaaaaa aaaaaaaacc cccccccccc ccccctgcag gcgcgccgcc ggcgatttaa
     2641 attaattaat ctagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg
     2701 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2761 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2821 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2881 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2941 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3001 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3421 ccggtaagac acgaccttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3841 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3901 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3961 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4021 tccagattta tcagcaataa accagccagc ccggaagggc cgagcgcaga agtggtcctg
     4081 caaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4621 tcagcatctt ttactttcac cagcgtttct gggtgagccc tttcaccagc gtttctgggt
     4681 gagc
//
