LOCUS       Exported                6634 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 6634)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6634
                     /organism="synthetic DNA construct"
                     /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..2784
                     /codon_start=1
                     /label=OposCharlie2
                     /translation="MENPKNKKTRLSKGSGSSSSRPFQETWTEMYGIIEKNGRSFCVLC
                     TETVVSRTWNINRHFETNHSQLLKKSEDERKEYISRQLHLYKSQSNSILKFVKGSTNLT
                     SASLSIAHSIAQHGKALSEGEFIKETLLRCAPVLFHDMQNKDAIIKRISELPLSRNIIK
                     DQIMRLNTNVQHQLKRDISNCKYFSISLDETTDVTSHAQLAIIGRYSDGLTMREELIKL
                     VSVPTSTSGSEICKVVIQTFRDLSIDISKVVSVTTDGAPNMVGKKVGFVKLFTEAIGHP
                     IVPFHCIIHQEALCAKAGFTDLNDLMSVVTKIVNLIAARPLHKREFSALLLEVDSTYSG
                     LLMYNNVRWLSRGKVLERFVECFEEIKVFLEDKDLGNFPQLNDDKWVNTLMFFTDLSVH
                     INELNLKLQGFGKSIDVMFGYIKAFESKLKIFKRDVETKTYKYFPRVTKYFEKASAAVQ
                     NEMEPLHIKYQHVLDSLLDQFSDRFNQFRSLEQTMKIIKYPDVVVYSSLELNGFQWMQI
                     DDLEMQLAEFQDSIWAQVFVDLRSKLENLERCRLENQEECHYEQEIWSAWNRLPDTFST
                     LKNIAMALLTIFPSTYFCETLFSALNNIKTNKRNRLTDEVSSACLGLKCTKYQPSIEDL
                     ANEIQQQKSH"
     promoter        complement(2822..2840)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2866..3090
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3136..3564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3578..3907
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3758..3893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4073..4194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4243..4259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4267..4283
                     /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(4291..4321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4336..4357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4645..5233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5404..6264)
                     /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(6265..6369)
                     /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 gccaccatgg agaaccccaa gaacaaaaaa acccgcctga gcaaaggttc aggtagcagc
      901 agcagccgtc cttttcagga aacgtggacc gaaatgtatg gcatcatcga aaaaaacggc
      961 cgcagcttct gtgttctgtg caccgaaacc gttgttagcc gcacctggaa catcaaccgc
     1021 catttcgaaa ccaaccatag ccagctgctg aagaaaagcg aagatgagcg caaagaatac
     1081 attagccgcc agctgcacct gtataagagc cagagcaaca gcattctgaa atttgtgaag
     1141 ggcagcacca atttaaccag cgccagctta agcattgccc atagcattgc gcagcacggc
     1201 aaagcattaa gcgaaggcga gttcattaaa gaaaccctgc tgcgctgtgc acctgttctg
     1261 ttccacgaca tgcagaacaa agatgcgatt attaagcgca tcagcgaatt acctctgagc
     1321 cgcaacatta tcaaggacca gatcatgcgc ctgaacacca acgtgcagca tcagctgaaa
     1381 cgcgatatta gcaactgcaa atactttagc atcagcctgg atgaaaccac cgatgtgacc
     1441 agccatgcgc agctggcgat tattggccgc tatagcgatg gcctgaccat gcgcgaagaa
     1501 ctgattaaac tggtgagcgt gcctactagc actagcggca gcgaaatttg caaggtggtg
     1561 attcagacct ttcgcgatct gagcattgat atcagcaaag tggtgagcgt gaccactgat
     1621 ggtgcgccta acatggtggg caaaaaagtg ggcttcgtga aactgtttac cgaagcgatc
     1681 ggccatccga tagtgccgtt tcattgcatt attcatcagg aagcgctgtg tgcgaaagcg
     1741 ggctttaccg atctgaacga tctgatgagc gtggtgacca aaattgtgaa cctgattgcg
     1801 gcacgccctc tgcataaacg cgaatttagc gcgctgctgc tggaagtgga tagcacctat
     1861 agcggcctgc tgatgtacaa caacgtgcgc tggctgagcc gtggtaaagt gctggaacgc
     1921 tttgtggaat gctttgagga gatcaaagtg ttcctggaag acaaagacct gggcaacttt
     1981 cctcagctga acgatgataa atgggtgaac accctgatgt tcttcaccga tctgagcgtg
     2041 cacattaacg agctgaacct gaagctgcag ggctttggca agagcattga tgtgatgttc
     2101 ggctacatca aagcgtttga aagcaagctg aagatcttta agcgcgatgt ggaaaccaaa
     2161 acctacaaat atttccctcg cgtgaccaaa tactttgaaa aagcgagcgc cgcggtgcaa
     2221 aacgagatgg aacctctgca tatcaaatac cagcatgtgc tggatagcct gctggatcag
     2281 tttagcgatc gctttaacca gtttcgcagc ctggaacaga ccatgaagat catcaagtat
     2341 cctgacgtgg tggtgtatag cagcctggag ctgaacggct ttcagtggat gcagatcgac
     2401 gatctggaaa tgcagctggc ggagttccag gatagcattt gggcgcaggt gtttgttgat
     2461 ctgcgcagca aactggagaa cttagagcgc tgccgcctgg aaaaccagga agaatgccat
     2521 tatgagcagg aaatttggag cgcgtggaac cgtctgccgg atacctttag caccctgaaa
     2581 aacattgcga tggcgctgct gaccattttc cctagcacct acttctgcga aaccctgttt
     2641 agcgcgctga acaacatcaa aaccaacaag cgcaaccgcc tgaccgatga agttagcagc
     2701 gcgtgtttag gcctgaagtg caccaaatat cagcctagca tcgaagatct ggccaacgaa
     2761 atccagcagc agaaaagcca ttgatgagcg gccgctcgag catgcatcta gagggcccta
     2821 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     2881 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2941 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3001 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3061 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3121 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3481 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3541 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3661 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3721 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3781 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3841 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3901 ttgcaaaaag ctccccgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3961 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4021 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4201 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5281 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5341 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5401 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5461 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5521 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5581 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5641 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5701 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5761 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5821 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5881 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5941 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6001 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6061 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6121 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6181 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6241 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6301 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6361 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6421 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6481 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     6541 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     6601 ctgcttcgcg atgtacgggc cagatatacg cgtt
//
