LOCUS       62056_11405        5254 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5254)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           41..83
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             173..880
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             917..2302
                     /codon_start=1
                     /label=TRIM21
                     /note="mouse RING E3 ubiquitin ligase that can be used
                     together with antibodies to degrade intracellular proteins 
                     (Clift et al., 2018)"
                     /translation="MSLEKMWEEVTCSICLDPMVEPMSIECGHCFCKECIFEVGKNGGS
                     SCPECRQQFLLRNLRPNRHIANMVENLKQIAQNTKKSTQETHCMKHGEKLHLFCEEDGQ
                     ALCWVCAQSGKHRDHTRVPIEEAAKVYQEKIHVALEKLRKGKELAEKMEMDLTMQRTDW
                     KRNIDTQKSRIHAEFALQNSLLAQEEQRQLQRLEKDQREYLRLLGKKEAELAEKNQALQ
                     ELISELERRIRGSELELLQEVRIILERSGSWNLDTLDIDAPDLTSTCPVPGRKKMLRTC
                     WVHITLDRNTANSWLIISKDRRQVRMGDTHQNVSDNKERFSNYPMVLGAQRFSSGKMYW
                     EVDVTQKEAWDLGVCRDSVQRKGQFSLSPENGFWTIWLWQDSYEAGTSPQTTLHIQVPP
                     CQIGIFVDYEAGVVSFYNITDHGSLIYTFSECVFAGPLRPFFNVGFNYSGGNAAPLKLC
                     PLKM"
     3'UTR           2349..2474
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus
                     laevis"
     promoter        complement(2579..2597)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2615..2631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2639..2655)
                     /label=lac operator
                     /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(2663..2693)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2708..2729)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3017..3605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3779..4636)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4637..4741)
                     /label=AmpR promoter
     primer_bind     5215..5231
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5238..5254
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg cgaattaatt cgagctcggt acccagcttg cttgttcttt ttgcagaagc
       61 tcagaataaa cgctcaactt tggcagatca attccccggt gctagcatga agcggcccgc
      121 tgctactaag aaggctggtc aggctaagaa gaagaagcta ccggtcgcca ccatggtgag
      181 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
      241 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
      301 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
      361 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
      421 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
      481 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
      541 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
      601 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
      661 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
      721 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
      781 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
      841 gggccgccac tccaccggcg gcatggacga gctgtacaag tccggactca gatctcgagc
      901 tcaagcttcg aattcaatgt ctctggaaaa gatgtgggag gaggtcacct gttctatctg
      961 cctggatccc atggtggagc ctatgagtat cgaatgtggc cattgctttt gcaaggaatg
     1021 catttttgaa gttgggaaga atgggggcag ttcatgtccc gagtgccggc aacagtttct
     1081 gctccgaaac ctcaggccca atagacatat agccaacatg gtggaaaacc ttaaacagat
     1141 agcccagaat accaagaaga gtacccagga aacgcactgc atgaagcatg gagagaagct
     1201 tcacctattc tgtgaggaag atgggcaggc cctttgctgg gtgtgtgccc agtctgggaa
     1261 acaccgggac cacaccaggg tccctattga agaggctgct aaggtatacc aggagaagat
     1321 ccacgtggct ttagaaaaac tgagaaaggg gaaagagttg gccgagaaga tggaaatgga
     1381 tctcacgatg caaagaacag actggaagag gaacattgac acccagaagt cgaggattca
     1441 cgcagagttc gcacttcaga atagcttgct ggctcaggag gagcagaggc agctgcagag
     1501 gctggagaag gatcaaaggg agtacctgag actcctgggg aagaaggagg ctgagctggc
     1561 tgagaagaac caggccctgc aggagctgat ctcagagctg gagaggagga ttcgtggttc
     1621 agagctggag ctactgcagg aggtgaggat catcctggaa aggagtggat cctggaacct
     1681 ggacacgtta gatattgacg ccccagacct aacaagcaca tgccctgtgc cagggcggaa
     1741 gaagatgctg aggacgtgtt gggttcatat tactctggat cgcaacaccg ccaactcatg
     1801 gctcatcatc tcaaaggatc ggagacaagt gaggatggga gacacccatc agaacgtgtc
     1861 tgacaataag gagaggttta gtaattaccc catggtgcta ggtgcccaga gattctcctc
     1921 tgggaagatg tactgggagg tagatgtgac tcaaaaggag gcctgggatc tgggggtttg
     1981 cagagattct gttcagagga aagggcagtt ttcactcagt cccgagaatg gcttctggac
     2041 catttggtta tggcaagaca gctatgaggc tggtaccagt cctcagacca ccctccacat
     2101 tcaagtacct ccatgccaaa ttgggatctt tgtggactat gaggctggcg ttgtctcctt
     2161 ctacaacata actgaccatg gctccctcat ttacaccttc tcggagtgtg tttttgctgg
     2221 acctctgcga cctttcttca atgttggttt caattatagt gggggaaatg cagcgcctct
     2281 aaagctctgt ccactaaaga tgtgagcggc cgctctagat aaatgatagc ttgatctggt
     2341 taccactaaa ccagcctcaa gaacacccga atggagtctc taagctacat aataccaact
     2401 tacactttac aaaatgttgt cccccaaaat gtagccattc gtatctgctc ctaataaaaa
     2461 gaaagtttct tcacattcta aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaccccccc
     2521 cccccccccc tgcaggcgcg ccgccggcga tttaaattaa ttaatctagc ttgagtattc
     2581 tatagtgtca cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2641 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2701 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2761 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2821 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4081 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4381 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4741 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4801 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4861 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4921 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4981 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5041 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     5101 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     5161 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     5221 cgacggccag tgaattgtaa tacgactcac tata
//
