LOCUS       pBS-SKII-3XHA-HA        5424 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 5424)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5424
                     /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..1745
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     promoter        1803..2146
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2147..2964
                     /codon_start=1
                     /label=C. alb. URA3
                     /translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
                     FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
                     KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
                     AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
                     TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRDAGWNAYLKKTGQL*S"
     terminator      2965..3162
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(3236..3254)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3275..3291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3299..3315
                     /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(3323..3353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3368..3389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3677..4265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4436..5296)
                     /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(5297..5401)
                     /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 aggatcctat ccatatgacg ttccagatta cgctgctcag tgcggttcag
      841 gtggatctgg ttctatggca gaaatcggta ctggctttcc attcgacccg cattatgtgg
      901 aagtcctggg cgagagaatg cattacgttg acgtgggtcc gagagatgga actccggtcc
      961 tttttctgca cgggaatcct acaagctctt atgtttggcg caatatcatc cctcatgtag
     1021 ctccgacgca tcgctgtatt gcgccggacc tgattggtat gggaaaatct gataaaccag
     1081 acctgggtta ctttttcgat gatcatgtgc gtttcatgga tgccttcatt gaggcattag
     1141 ggcttgaaga agtcgtcttg gtgattcatg attggggctc agctctggga tttcactggg
     1201 ctaaaagaaa tcctgaacgc gtaaaaggca tcgcgtttat ggagttcatt cgtccaattc
     1261 cgacttggga tgaatggcct gagttcgcga gagaaacatt tcaagcattt cgcacgaccg
     1321 atgtaggccg gaagttaatc atcgatcaga atgtctttat cgaagggaca ttgccgatgg
     1381 gagtcgttcg tccgttaaca gaagtcgaaa tggatcacta tagagaacct tttcttaatc
     1441 ctgtggacag agagccgctg tggcggtttc cgaacgaact gccgattgca ggcgagcctg
     1501 ctaacattgt agcgctggtt gaagagtata tggattggct tcatcagtct ccagttccga
     1561 agttattgtt ttggggtacg cctggcgtgc ttattccacc ggccgaagcg gcacgtttgg
     1621 caaaaagcct gccaaattgc aaagccgttg acattggccc tggacttaac ttgcttcaag
     1681 aggataaccc ggacttaatc gggagcgaaa ttgcccggtg gctttctacc ttagaaatca
     1741 gcggctaata tataactgtc tagaaataaa gagtatcatc tttcaaaggt cacccggcca
     1801 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     1861 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     1921 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg
     1981 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     2041 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     2101 tcttgttagg atacagttct cacatcacat ccgaacataa acaaccatga cagtcaacac
     2161 taagacctat agtgagagag cagaaactca tgcctcacca gtagcacaac gattatttcg
     2221 attaatggaa ctgaagaaaa ccaatttatg tgcatcaatt gatgttgata ccactaagga
     2281 attccttgaa ttaattgata aattgggtcc ttatgtatgc ttaatcaaga ctcatattga
     2341 tataatcaat gatttttcct atgaatccac tattgaacca ttattagaac tttcacgtaa
     2401 acatcaattt atgatttttg aagatagaaa atttgctgat attggtaata ccgtgaagaa
     2461 acaatatatt ggtggagttt ataaaattag tagttgggca gatattacta atgctcatgg
     2521 tgtcactggg aatggagtag ttgaaggatt aaaacaggga gctaaagaaa ccaccaccaa
     2581 ccaagagcca agagggttat tgatgttagc tgaattatca tcagtgggat cattagcata
     2641 tggagaatat tctcaaaaaa ctgttgaaat tgctaaatcc gataaggaat ttgttattgg
     2701 atttattgcc caacgtgata tgggtggaca agaagaagga tttgattggc ttattatgac
     2761 acctggagtt ggattagatg ataaaggtga tggattagga caacaatata gaactgttga
     2821 tgaagttgtt agcactggaa ctgatattat cattgttggt agaggattgt ttggtaaagg
     2881 aagagatcca gatattgaag gtaaaaggta tagagatgct ggttggaatg cttatttgaa
     2941 aaagactggc caattataat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     3001 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     3061 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     3121 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     3181 cgccgccact agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt
     3241 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3301 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3361 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3421 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3481 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4081 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5401 gcacatttcc ccgaaaagtg ccac
//
