LOCUS       Exported                6508 bp ds-DNA     circular SYN 22-FEB-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6508)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6508
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             847..2658
                     /codon_start=1
                     /label=HAT1_AG
                     /translation="MMAPTNATTSPVWDHFSPVETGAKCLYCLKVFKYTKGTTSNLKRH
                     LNLVHKTVPYLKQKQPIPQTINIDDEAGPSAVNFQPSNQYFNSNMSIQGYLKKPINSET
                     KKVLDRMLLDLICKECLPFNLVESEIFKKFVYTLNPNYIMPTRKSLSNALLPSVYNQEF
                     EKAKEKLSTAKAIAITSDGWTNLNQISFFALTGHYIDENCKLSSILIECSEFENPHSGR
                     NIANWIQGTLNKFDIEDKIVAMVTDNASNMKAASTELNFCHIPCFAHTLNLIVRDAIKK
                     SVLPVVEEVKRVVMLFKKSPKASQMLADTQKKLNLDQLKMIQEVSTRWNSGYDMLNRFY
                     KNKIALLSCADSLKMKISLESHDWEAIEQIVRVLKYFYSATNIVSAQKYITISHVGLLC
                     NVLLTKTSQFRNDEDIAENIQNLVALLIEGLQNKLKIYRSNEQILKSMILDPRIKQLGF
                     QDDVEKFKNICESIISELLPLQKPAVEVEKVVKKVSKDVDMLFGDLLKNKGAQNYKTPR
                     QIAENELHQYLSVENIDLENDPLLWWKEHQVLYPSLYTLAMSTLCIPGTSVPCERLFSK
                     AGQIYSEKRSRLAPKKLQEILFIQQNA"
     CDS             1642..1644
                     /codon_start=1
                     /label=C265A C266A TGC-GCC
                     /translation="C"
     CDS             1651..1653
                     /codon_start=1
                     /label=H268A H269A CAT-GCT
                     /translation="H"
     CDS             1663..1665
                     /codon_start=1
                     /label=L272A L273A CTG-GCG
                     /translation="L"
     CDS             1828..1830
                     /codon_start=1
                     /label=R318A R328A CGT-GCT
                     /translation="R"
     CDS             1837..1839
                     /codon_start=1
                     /label=S321A S331A AGC-GCC
                     /translation="S"
     CDS             2197..2199
                     /codon_start=1
                     /label=P437A P451A CCG-GCG
                     /translation="P"
     CDS             2569..2571
                     /codon_start=1
                     /label=S576A S575A AGC-GCC
                     /translation="S"
     CDS             2599..2601
                     /codon_start=1
                     /label=R586A R585A CGC-GCC
                     /translation="R"
     promoter        complement(2696..2714)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2740..2964
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3010..3438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3452..3781
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3632..3767
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3947..4068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4141..4157
                     /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(4165..4195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4210..4231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4519..5107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5278..6138)
                     /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(6139..6243)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacgtc cgcgcagggc aatagcagag ctctctggct
      721 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      781 ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcgcc
      841 gccaccatga tggcgcctac caacgcaacc actagccctg tgtgggatca ttttagcccg
      901 gtggaaaccg gcgcgaaatg cctgtattgc ctgaaggtgt tcaaatatac caaaggcacc
      961 accagcaact taaaacgcca cctgaacctg gtgcacaaaa ccgtgcctta tctgaaacag
     1021 aagcagccta ttccgcagac cattaacatt gatgatgagg cgggtccgag cgcggttaat
     1081 tttcagccga gcaaccagta ttttaacagc aacatgagca ttcagggcta tctgaagaag
     1141 cctattaaca gcgaaaccaa aaaggtgctg gaccgcatgc tgctggatct gatctgcaag
     1201 gagtgcctgc cttttaacct ggtggaaagc gagattttta agaagttcgt gtacaccctg
     1261 aaccctaact atatcatgcc gacccgcaaa tcactgagca acgcgctgtt acctagcgtg
     1321 tacaaccagg aatttgagaa agcgaaggaa aaactgagca ccgcgaaagc gatcgccatt
     1381 acctcagatg gctggaccaa cctgaaccag attagctttt ttgcgctgac cggccattat
     1441 attgacgaaa actgcaaact gagcagcatc ctgattgaat gcagcgaatt tgagaacccg
     1501 catagcggcc gtaatatcgc gaactggatt cagggcaccc tgaacaaatt tgacatcgag
     1561 gataagattg tggcgatggt gaccgataac gcgagcaaca tgaaagcagc gagcaccgag
     1621 ctgaactttt gccatatccc gtgctttgcg cataccctga acctgattgt gcgcgatgcg
     1681 attaagaaaa gcgtgctgcc tgtggtggag gaagtgaaac gcgtggtgat gctgtttaaa
     1741 aaaagcccga aagcgagcca aatgctggcg gatacccaga aaaaactgaa cctggatcag
     1801 ctgaagatga ttcaggaagt gagcacccgt tggaacagcg gctatgacat gctgaaccgc
     1861 ttttacaaga acaaaattgc cctgctgagc tgtgcagaca gcctgaaaat gaaaatcagc
     1921 ctggagagcc atgattggga agcgatcgaa cagatcgtgc gcgtgctgaa atacttttat
     1981 agcgcgacca acatcgtgag cgcgcagaaa tatatcacca ttagccatgt gggcctgtta
     2041 tgcaacgtgc tgctgaccaa aaccagccag tttcgcaacg atgaggacat tgccgaaaac
     2101 attcagaacc tggtggccct gctgatagaa ggcctgcaga acaaactgaa aatctaccgc
     2161 agcaacgaac agattctgaa gagcatgatt ctggacccgc gcattaaaca gctgggcttt
     2221 caggacgatg tggagaagtt caaaaacatc tgcgagagca ttattagcga actgctgcct
     2281 ctgcagaaac ctgcggtgga agtggaaaag gtggtgaaaa aagtgagcaa ggacgtggac
     2341 atgctgtttg gcgatctgct gaagaacaaa ggcgcccaga actataaaac cccgcgccag
     2401 atcgcagaga acgaactgca tcagtacctg agcgtggaaa acattgatct ggaaaacgat
     2461 ccgctgctgt ggtggaaaga acaccaggtg ctgtatccta gcctgtatac cctggcgatg
     2521 agcaccctgt gtattcctgg caccagcgtt ccttgcgaac gcctgtttag caaagcgggc
     2581 cagatctata gcgaaaaacg cagccgcctg gcgcctaaaa agctgcagga aatcctgttt
     2641 atccagcaga acgcgtgatg agcggccgct cgagcatgca tctagagggc cctattctat
     2701 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     2761 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2821 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2881 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2941 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3001 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3061 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3121 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3181 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3241 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3301 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3361 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3421 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3481 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3601 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3661 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3721 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3781 aaagctcccc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3841 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3901 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4081 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4921 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6241 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6301 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6361 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6421 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     6481 cgcgatgtac gggccagata tacgcgtt
//
