LOCUS       pBS-SKII-3XHA-fS        5078 bp DNA     circular SYN 20-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5078)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             735..824
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             855..1400
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     promoter        1458..1801
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1802..2619
                     /codon_start=1
                     /label=C. alb. URA3
                     /translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
                     FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
                     KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
                     AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
                     TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRDAGWNAYLKKTGQL*S"
     terminator      2620..2817
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(2890..2908)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2929..2945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2953..2969
                     /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)."
     promoter        complement(2977..3007)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3022..3043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4090..4950)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4951..5055)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gggtcgacgg atccccgggt
      721 taattaacat cttttaccca tacgatgttc ctgactatgc gggctatccc tatgacgtcc
      781 cggactatgc gggatcctat ccatatgacg ttccagatta cgctgctcag tgcggttcag
      841 gtggatctgg ttctatggac aaagactgcg aaatgaagcg caccaccctg gatagccctc
      901 tgggcaagct ggaactgtct gggtgcgaac agggcctgca ccgtatcatc ttcctgggca
      961 aaggaacatc tgccgccgac gccgtggaag tgcctgcccc agccgccgtg ctgggcggac
     1021 cagagccact gatgcaggcc accgcctggc tcaacgccta ctttcaccag cctgaggcca
     1081 tcgaggagtt ccctgtgcca gccctgcacc acccagtgtt ccagcaggag agctttaccc
     1141 gccaggtgct gtggaaactg ctgaaagtgg tgaagttcgg agaggtcatc agctacagcc
     1201 acctggccgc cctggccggc aatcccgccg ccaccgccgc cgtgaaaacc gccctgagcg
     1261 gaaatcccgt gcccattctg atcccctgcc accgggtggt gcagggcgac ctggacgtgg
     1321 ggggctacga gggcgggctc gccgtgaaag agtggctgct ggcccacgag ggccacagac
     1381 tgggcaagcc tgggctgggt taatatataa ctgtctagaa ataaagagta tcatctttca
     1441 aaggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc
     1501 gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat
     1561 aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct
     1621 cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg
     1681 ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata
     1741 tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac
     1801 catgacagtc aacactaaga cctatagtga gagagcagaa actcatgcct caccagtagc
     1861 acaacgatta tttcgattaa tggaactgaa gaaaaccaat ttatgtgcat caattgatgt
     1921 tgataccact aaggaattcc ttgaattaat tgataaattg ggtccttatg tatgcttaat
     1981 caagactcat attgatataa tcaatgattt ttcctatgaa tccactattg aaccattatt
     2041 agaactttca cgtaaacatc aatttatgat ttttgaagat agaaaatttg ctgatattgg
     2101 taataccgtg aagaaacaat atattggtgg agtttataaa attagtagtt gggcagatat
     2161 tactaatgct catggtgtca ctgggaatgg agtagttgaa ggattaaaac agggagctaa
     2221 agaaaccacc accaaccaag agccaagagg gttattgatg ttagctgaat tatcatcagt
     2281 gggatcatta gcatatggag aatattctca aaaaactgtt gaaattgcta aatccgataa
     2341 ggaatttgtt attggattta ttgcccaacg tgatatgggt ggacaagaag aaggatttga
     2401 ttggcttatt atgacacctg gagttggatt agatgataaa ggtgatggat taggacaaca
     2461 atatagaact gttgatgaag ttgttagcac tggaactgat attatcattg ttggtagagg
     2521 attgtttggt aaaggaagag atccagatat tgaaggtaaa aggtatagag atgctggttg
     2581 gaatgcttat ttgaaaaaga ctggccaatt ataatcagta ctgacaataa aaagattctt
     2641 gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca
     2701 aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta
     2761 agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg
     2821 tcgattcgat actaacgcgc cactagttct agagcggccg ccaccgcggt ggagctccag
     2881 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     2941 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3001 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3061 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5041 aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
