LOCUS       Exported                5719 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 5719)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5719
                     /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..1869
                     /codon_start=1
                     /label=Tc1-1_Xt
                     /translation="MPRSKEIQEQMRTKVIEIYQSGKGYKAISKALGLQRTTVRAIIHK
                     WQKHGTVVNLPRSGRPTKITPRAQRQLIREATKDPRTTSKELQASLASIKVSVHDSTIR
                     KRLGKNGLHGRFPRRKPLLSKKNIKARLNFAKKHLNDCQDFWENTLWTDETKVELFGRC
                     VSRYIWRKSNTAFQKKNIIPTVKYGGGSVMVWGCFAASGPGRLAVIDGTMNSTVYQKIL
                     KENVRPSVRQLKLKRSWVLQQDNDPKHTSKSTSEWLKKNKMKTLEWPSQSPDLNPIEML
                     WHDLKKAVHARKPSNKAELQQFCKDEWAKIPPERCKRLVASYRKRLIAVIAAKGGPTSY
                     "
     CDS             1591..1593
                     /codon_start=1
                     /label=aH249A CAC-GCC
                     /translation="H"
     CDS             1687..1689
                     /codon_start=1
                     /label=aL281A CTG-GCG aL281V CTG-GTG
                     /translation="L"
     CDS             1690..1692
                     /codon_start=1
                     /label=aW282F TGG-TTT
                     /translation="W"
     CDS             1744..1746
                     /codon_start=1
                     /label=aL300A CTG-GCG aL300T CTG-ACG
                     /translation="L"
     CDS             1822..1824
                     /codon_start=1
                     /label=R326H CGC-CAC
                     /translation="R"
     polyA_signal    1951..2175
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2221..2649
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2663..2992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2843..2978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3158..3279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3328..3344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3352..3368
                     /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(3376..3406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3421..3442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3730..4318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4489..5349)
                     /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(5350..5454)
                     /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 gccaccatgc ctcgcagcaa agagattcag gaacagatgc gcaccaaggt gattgaaatc
      901 tatcagagcg gcaaaggcta taaagcgatt agcaaggccc tgggcttaca acgtactacc
      961 gtgcgcgcca ttattcacaa atggcagaaa catggcaccg tggttaattt acctcgcagc
     1021 ggtcgtccta ccaaaattac ccctcgcgcg cagcgtcaat taattcgtga agccaccaag
     1081 gatcctcgta ccaccagcaa agaactgcaa gccagcctgg ccagcataaa agtgagcgtg
     1141 catgatagca ccattcgcaa gcgcttaggc aagaatggtc tgcacggccg ttttcctcgt
     1201 cgtaaaccgc tgctgagcaa gaaaaacatt aaagcgcgcc tgaactttgc gaaaaagcat
     1261 ctgaacgatt gccaggactt ttgggaaaac accctgtgga ccgatgaaac caaggtggaa
     1321 ctgttcggtc gctgcgtgag ccgttatatc tggcgcaaaa gcaacaccgc gttccagaag
     1381 aagaacatta ttccgaccgt gaagtatggc ggcggtagcg ttatggtgtg gggttgtttt
     1441 gcggcatcag gccctggtcg tttagcagtt atcgatggca ccatgaacag caccgtgtac
     1501 cagaaaatcc tgaaagaaaa cgtgcgtccg agcgtgcgcc agttaaaact gaaacgcagc
     1561 tgggtgctgc aacaggacaa cgatccgaaa cacaccagca aaagcaccag cgagtggctg
     1621 aagaagaaca agatgaaaac cctggagtgg cctagccaaa gccctgatct gaacccgatc
     1681 gaaatgctgt ggcacgatct gaaaaaagcc gtgcatgcgc gcaaacctag caacaaagcg
     1741 gagctgcagc agttctgcaa agacgaatgg gcgaaaattc cgccggaacg ctgtaaacgt
     1801 ctggtggcca gctatcgcaa acgcctgata gccgtgatcg cagcaaaagg tggtcctacc
     1861 agctactaat gagcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     1921 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1981 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2041 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2101 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2161 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2221 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2281 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2341 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2401 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2461 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2521 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2581 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2641 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2761 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2821 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2881 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2941 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3001 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3061 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3121 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     3181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     3301 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4141 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5461 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     5521 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     5581 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     5641 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     5701 cgggccagat atacgcgtt
//
