LOCUS       pCfB2197-PrGNP1-        7622 bp 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 7622)
  AUTHORS   RNAcious laboratory
  TITLE     Direct Submission
  JOURNAL   Exported Feb 22, 2024 from SnapGene 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..201
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    249..282
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            337..1456
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        337..680
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             681..1253
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1259..1456
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1519..1552
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(1596..1614)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1624..1640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2332..2365)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(2835..2851)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3121..3709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3833..4624,4625..4693))
                     /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(4694..4785)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    5078..5111
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     terminator      complement(5704..5869)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        5909..6908
                     /label=GNP1 promoter
     gene            6909..7622
                     /label=mKate2
ORIGIN
        1 cgcgtgcatt catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
       61 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
      121 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
      181 aaggttttgg gacgctcgaa gatcgcgtca gctgaagctt cgtacgctgc aggtcgacaa
      241 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtcta gagatctgtt
      301 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
      361 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
      421 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
      481 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
      541 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
      601 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
      661 acatccgaac ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca
      721 ccagtgtccc gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca
      781 ccgttttccg cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc
      841 cgcccctgac caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg
      901 acggcgaccc ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct
      961 tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg
     1021 ccccggagca ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg
     1081 cccgcgagcg gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga
     1141 tccacgcgta ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg
     1201 gcaccgcctc ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac
     1261 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     1321 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     1381 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     1441 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag
     1501 ctctcgagaa cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtgat
     1561 atcagatcca ctagtggcct atgcacccaa ttcgccctat agtgagtcgt attacgcgcg
     1621 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccctgcagg
     1681 actagtgctg aggcattaat cttcccgtga atcaactgca cataccttaa taactagttc
     1741 agtttagtgc tctgtctgag tgactgtaat aataaatttt acaagccact tctcatgaca
     1801 tatattggta agtaacttca tcaatactaa ttagtctttg ccggttaccc atctggcccc
     1861 tgacttgcga tgcttaggaa gttccatact cgcggctctt cccaacagta gcacatccgt
     1921 gaaacttctg gcgctattca ttatgcagta ccaggacaag aagttaaaaa aaaagctctg
     1981 ttacaagttc aatggtggtg caaggattga agttattatc caggaggcac ggatgcaaaa
     2041 gatagaggac aaattaattt ccttaaaatt caaaatgctc attattgtca cgctgtatga
     2101 gcatttggtg aagatttcac tgggaaatgt tgcaataatt tgataatcgt tcgtattgga
     2161 tgaaactgta acatcatctg tttattaagt atccgtgtta ttagtatatc atcacatacg
     2221 gtgtaagaag ataacataaa gattgagaaa cagtcatcaa atataatgga agctgaaatg
     2281 cgaggattga ttaagacctc agcgcggccg caaatttaaa taaaatgaag tgaagttcct
     2341 atactttcta gagaatagga acttctatag tgagtcgaat aagggcgaca caaaatttat
     2401 tctaaatgca taataaatac tgataacatc ttatagtttg tattatattt tgtattatcg
     2461 ttgacatgta taattttgat atcaaaaact gattttccct ttattatttt cgagatttat
     2521 tttcttaatt ctctttaaca aactagaaat attgtatata caaaaaatca taaataatag
     2581 atgaatagtt taattatagg tgttcatcaa tcgaaaaagc aacgtatctt atttaaagtg
     2641 cgttgctttt ttctcattta taaggttaaa taattctcat atatcaagca aagtgacagg
     2701 cgcccttaaa tattctgaca aatgctcttt ccctaaactc cccccataaa aaaacccgcc
     2761 gaagcgggtt tttacgttat ttgcggatta acgattactc gttatcagaa ccgcccaggg
     2821 ggcccgagct taagactggc cgtcgtttta caacacagaa agagtttgta gaaacgcaaa
     2881 aaggccatcc gtcaggggcc ttctgcttag tttgatgcct ggcagttccc tactctcgcc
     2941 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3001 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3061 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3121 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3181 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3241 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3301 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3361 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3421 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3481 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtgggc
     3541 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3601 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3661 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3721 gatcttttct acggggtctg acgctcagtg gaacgacgcg cgcgtaactc acgttaaggg
     3781 attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc ttttaccaat
     3841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagcgctg
     3961 cgatgatacc gcgagaacca cgctcaccgg ctccggattt atcagcaata aaccagccag
     4021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4141 ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4441 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4681 gttgaatact catattcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gtcagtgtta
     4801 caaccaatta accaattctg aacattatcg cgagcccatt tatacctgaa tatggctcat
     4861 aacacccctt gtttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     4921 tcagaagtga aacgccgtag cgccgatggt agtgtgggga ctccccatgc gagagtaggg
     4981 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgcccggg
     5041 ctaattatgg ggtgtcgccc ttattcgact ctatagtgaa gttcctattc tctagaaagt
     5101 ataggaactt ctgaagtggg gatttaaatg cggccgcgct gagggtttaa tgttgagctc
     5161 tgtccttcat ggactttttg gaccacttct tcttggactt cttaccacca gcaagagcgg
     5221 cagcggcttt ggcagcttta gacaattgtt gctttggagg catgttatat tatgttctga
     5281 gacgtaagaa agggtgaaaa ttgatgttag tgtcaaaaat tatattacaa aatacgcaga
     5341 gatattctag ttcctttgat gaatgaatct ttcagaaaaa aaagtcaaag caaaagcaaa
     5401 atggcctgca gactaaactg tatggtggtc ttggaatgat aaagatctgt ttaatagatt
     5461 tagtagatac aatagcacat ctcattaccc agttatgatt gacgtcattc tgagttacaa
     5521 tgatcttagc gacctgtgct cattatttgc tccactaatt ctaattttcc tcgcctttca
     5581 tatttcgtat ctttattcta tatcctaaaa tttttttggc aaatcccaga tttggctttg
     5641 attttggcat cggttcggtt ctttcattaa gtcctcagcg agctcgcatg gaatgcgtgc
     5701 gatgagcgac ctcatgctat acctgagaaa gcaacctgac ctacaggaaa gagttactca
     5761 agaataagaa ttttcgtttt aaaacctaag agtcacttta aaatttgtat acacttattt
     5821 tttttataac ttatttaata ataaaaatca taaatcataa gaaattcgct tatttagaag
     5881 tgtcaacaac gtatctacca acggaatgct tgacctctgg cttagcttct tgattgactt
     5941 ctgagtccga catcgctcgt gtatttattt gtaaaactag aactgaactg ggtccttctg
     6001 tcctatatgc tacaggtggt aacaagaaac gggagcttgg aaaaggtaag aaaaatcaac
     6061 taaaagagta gaatgtgttc gtttgtttga ttgttggtgt tttgctcttt tcctgcttcc
     6121 tgaacacctt ccgacggcgc gtgttggcaa agctgggtaa cccaacatat tcatatcaca
     6181 tgatcaaaga gttcggcggc aaaaacgtga catgcacgtg atggaaacag catggctttt
     6241 ttcagtttta tttttttttt tcactttcaa aagcattatt ccttggggac tacgtaatca
     6301 acgcgttggt catttctttt tccaacgcgc atgactaacc tactatgacg acgtagcctt
     6361 aattcggcat taccgtatta tgcgacgtgc ttgcgatcct tgaaaacggc atcaggatgc
     6421 acaaaaacgt ttcaaaagtg ccccgccgta cggtatgcgg cgccaaaaac cggtgaacat
     6481 gaaaaagaaa cggcacaggg accatcaaaa ataggaaaaa agaataacgc cgcatgaaaa
     6541 gaaagcggat ggaaaggcac ggcaactctt agagatgaag gcgatgatgc gatggcagac
     6601 ggacaatgaa atgtaaattt gaagtttttt tcggaaatcg gttttagtgt tcgtatgtca
     6661 gtttggctcg ggttgtgcag tgtcgtttct tttttgagtc ccttcttccc ccacctcttt
     6721 gatgttagaa agaatcggat tctgtatata taaccgattt ttttcacatt tctgttgatt
     6781 taaacatatt cctttcgtgc ttcttcaaag ttgagagtta taggtgcttt atgttttttt
     6841 ttttttgtgt aagaaaaaaa aactatcgct ttctcaagta gcttatataa tatcaaatat
     6901 tgcacattat gggttctggt ggttctgaat tgatcaaaga aaacatgcac atgaagttgt
     6961 acatggaagg tactgttaac aaccatcatt tcaagtgtac ctctgaaggt gaaggtaaac
     7021 catatgaagg tactcaaacc atgagaatca aagctgttga aggtggtcca ttgccatttg
     7081 cttttgatat tttggctacc tccttcatgt acggttctaa gactttcatt aaccacaccc
     7141 aaggtatccc agatttcttc aaacaatctt tcccagaagg tttcacttgg gaaagagtta
     7201 ctacttacga agatggtggt gttttgactg ctactcaaga tacttcttta caagacggtt
     7261 gcttgatcta caacgttaag attagaggtg tcaacttccc atctaatggt ccagttatgc
     7321 aaaaaaagac tttgggttgg gaagcttcta ctgaaacatt atatccagct gatggtggtt
     7381 tggaaggtag agctgatatg gctttgaaat tggttggtgg tggtcatttg atctgcaact
     7441 tgaaaactac ctacagatct aagaagccag ccaagaattt gaaaatgcca ggtgtttact
     7501 acgtcgatag aagattggaa agaatcaaag aagccgacaa agaaacctac gttgaacaac
     7561 acgaagttgc tgttgctaga tattgtgatt tgccatccaa attaggtcat agaggatctt
     7621 aa
//
