LOCUS       pBS-SKII-3XHA-HA        4998 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 4998)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4998
                     /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..2538
                     /codon_start=1
                     /label=PhleoR/BleR
                     /translation="MGMTDQATPNLPSRDFDPTAAFYERLGFGIVFRDAGWMILQRGDL
                     MLEFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGT
                     MAALVDPDGTLLRLIQNELLAGIS*S"
     terminator      2539..2736
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(2810..2828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2849..2865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2873..2889
                     /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(2897..2927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2942..2963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3251..3839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4010..4870)
                     /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(4871..4975)
                     /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 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg gtatgaccga
     2161 ccaagcgacg cccaacctgc catcacgaga tttcgatccc accgccgcct tctatgaaag
     2221 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     2281 catgctggag ttcttcgccc accccgggct cgatcccctc gcgagttggt tcagctgctg
     2341 cctgaggctg gacgacctcg cggagttcta ccggcagtgc aaatccgtcg gcatccagga
     2401 aaccagcagc ggctatccgc gcatccatgc ccccgaactg caggagtggg gaggcacgat
     2461 ggccgctttg gtcgacccgg acgggacgct cctgcgcctg atacagaacg aattgcttgc
     2521 aggcatctca tgatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     2581 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     2641 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     2701 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     2761 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     2821 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2881 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2941 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3001 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4981 ttccccgaaa agtgccac
//
