LOCUS       Exported                5611 bp DNA     circular SYN 03-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5611)
  AUTHORS   Matthew
  TITLE     Direct Submission
  JOURNAL   Exported Mar 3, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    80..104
                     /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)."
     misc_feature    142..145
                     /label=Probable TATA box
     misc_feature    148..178
                     /label=cLIC Cassette
     CDS             188..898
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins"
                     /label=Folding reporter GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             905..934
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             979..996
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1063..1110
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1147..1575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1906..2862)
                     /codon_start=1
                     /label=RepMyco
                     /translation="MAACAEGLRRAVDGDRSYSGLMTKNPGHIAWETEWLHSDLYTLSH
                     IEAELGANMPPPRWRQQTTYKAAPTPLGRNCALFDSVRLWAYLPALMRIYLPTRNVDGL
                     GRAIYAECHARNAEFPCNDVCPGPLPDSEVRAIANSIWRWITTKSRIWADGIVVYEATL
                     SARHAAISRKGAAARTAASTVARRAKSASAMHGGIAMSDGYSDGYSDGYNWQPTVRKKR
                     RVTAAEGARITGLSERHVVRLVAQERSEWFAEQAARRERIRAYHDDEGHSWPQTAKHFG
                     LHLDTVKRLGYRARKERAAEQEAAQKAHNEADNPPLF"
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /label=pBR322 ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             join(4972..5611,1..89)
                     /codon_start=1
                     /label=HygR
                     /translation="MSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGAGDAGHDAS
                     GVEDRDLDPAKLIRLTIGEL"
ORIGIN
        1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
       61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
      121 tgtttaactt taagaaggag atataccatg ggtggtggat ttaaatttat acttccaagg
      181 tcaattcagc aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      241 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgctacata
      301 cggaaaactc acccttaaat ttatttgcac tactggaaaa ctacctgttc cttggccaac
      361 acttgtcact actctgacct atggtgttca atgcttttcc cgttatccgg atcacatgaa
      421 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc
      481 tttcaaagat gacgggaact acaagacgcg tgctgaagtc aagtttgaag gtgataccct
      541 tgttaatcgt atcgagttaa agggtattga ttttaaagaa gatggaaaca ttctcggaca
      601 caaactagag tacaactata actcacacaa tgtatacatc acggcagaca aacaaaagaa
      661 tggaatcaaa gctaacttca aaattcgcca caacattgaa gatggttccg ttcaactagc
      721 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
      781 ttacctgtcg acacaatctg ccctttcgaa agatcccaac gaaaagcgtg accacatggt
      841 ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaaac
      901 tagtcatcat catcaccatc atcatcacca tcattaagtc gaccatctag agagctcaag
      961 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     1021 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1081 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     1141 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1501 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1561 aatttaacgc gaattttaac aaaatatcta gatttaaaga tctggtaccg cggccgcgcc
     1621 acggatgcca ccacaagcac tacaacggag ttcgccacgt acatcaccac aaccaccgat
     1681 tctggcggtg agctccacga tattcagcgg aaatggcttg gtatcgacca agattcgtag
     1741 aaccccgtct cgtctggctg gtattcaaaa cggacgcaac gaaacacgca acgagacagg
     1801 catggcccaa accagaaaac tagcgtctac caggactttt acctgtccga cccgttgcaa
     1861 cggaaccccc cacggaaccc ccgcgacacc cgctccccaa ttgcgttaga acagcggtgg
     1921 attgtcggct tcgttgtggg ccttttgagc cgcttcctgt tctgccgcac gctctttcct
     1981 cgcccgatag ccgagtcgct taacggtgtc cagatgcagc ccgaaatgtt tggccgtttg
     2041 cggccaagag tggccctcgt cgtcgtgata ggcgcggatg cgttcgcggc gtgcagcctg
     2101 ctcggcgaac cactcgctgc gttcctgcgc cacgagccgg acgacgtggc gttcggatag
     2161 tccggtgatt cgagcgcctt cggcggcggt cacgcgccgc tttttgcgga cagtcggctg
     2221 ccagttgtag ccgtcgctgt agccgtcgct gtagccgtcg ctcatagcaa tgcctccatg
     2281 catggctgac gcggactttg cgcgccgcgc aactgtgctc gccgccgtgc gcgctgctgc
     2341 gcccttccgc gagatggccg catggcgcgc actgagtgtg gcctcgtaga ccacgatccc
     2401 gtccgcccaa atgcgcgact tggttgtgat ccaacgccaa atgctgttgg cgatggcgcg
     2461 gacctcgctg tccggtagcg gtccgggaca cacgtcgttg cacggaaatt cggcgtttcg
     2521 cgcgtggcac tcggcataga tcgcgcggcc gagtccgtcc acgttccggg tcggcaggta
     2581 gatccgcatg agggcgggaa gataggccca caacctgacg gaatcgaaca gtgcgcaatt
     2641 ccgccctagc ggcgtcggag ccgctttgta cgtggtctgc tgacgccagc gcggcggtgg
     2701 catgttcgcg ccgagctcgg cctcgatgtg gctgagtgtg tagagatctg agtggagcca
     2761 ttccgtttcc caggcgatgt ggccggggtt tttggtcatg aggcctgagt aactgcggtc
     2821 gccatcgacg gcgcgccgaa ggccttcggc gcacgccgcc atgtatgcga gcggcttacg
     2881 ccgcgcgtat tcggtgcgtg gaacaggggc gttgagtgcc cacactgcgt gtgcgtggcc
     2941 gttggcgcga ttgcccacga tcgcgttggg cagcggatgg gacccccggg cgctgagcgc
     3001 tcggagcgct gcgtctggat ggtctacgtc cacgaccagc aggtttgcca gcgctgttgg
     3061 gttcgcctcg atgtaccggc ggcctagggc cgacgcgcgg ctttggcggt agatcccctc
     3121 gagcagatcg tcgcttgcca gggaccagta cggcagccag agctgctcaa attcgtcggc
     3181 gacgtggctc acgcttgata ataaaccacg attaatcacc ggtgtatggt ccgacacgag
     3241 ctccaagtca gatatttcgc tgaggggcca ccccacaact gcacactccc ccgctctccc
     3301 gtcgagccct gatgatgaaa caccagcgac agccgagcac ccccaaccac ctgtaccaac
     3361 caggaggaac acatgcgtcg tttcgaggac gtttccgggc cgctaagagc cgctgtggcg
     3421 gccgtacacg ccgccttaga cccgttagac cccctgccgc ctgaatgcgc gggtacgagc
     3481 cacacagcac ccgaacttac ggagctggtg ggctcacgcc gtgcggccgc acgcgttgcg
     3541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4321 gaactagagc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4501 ccccgacgtg gccgaccagc ccgtcatcgt caacgcctga ccgcggtgcg gacaggccgt
     4561 gtcgcgaccg gccgtgcgga attaagccgg cccgtaccct gtgaatagag gtccgctgtg
     4621 acacaagaat ccctgttact tctcgaccgt attgattcgg atgattccta cgcgagcctg
     4681 cggaacgacc aggaattctg ggagccgctg gcccgccgag ccctggagga gctcgggctg
     4741 ccggtgccgc cggtgctgcg ggtgcccggc gagagcacca accccgtact ggtcggcgag
     4801 cccgacccgg tgatcaagct gttcggcgag cactggtgcg gtccggagag cctcgcgtcg
     4861 gagtcggagg cgtacgcggt cctggcggac gccccggtgc cggtgccccg cctcctcggc
     4921 cgcggcgagc tgcggcccgg caccggagcc tggccgtggc cctacctggt gatgagccgg
     4981 atgaccggca ccacctggcg gtccgcgatg gacggcacga ccgaccggaa cgcgctgctc
     5041 gccctggccc gcgaactcgg ccgggtgctc ggccggctgc acagggtgcc gctgaccggg
     5101 aacaccgtgc tcacccccca ttccgaggtc ttcccggaac tgctgcggga acgccgcgcg
     5161 gcgaccgtcg aggaccaccg cgggtggggc tacctctcgc cccggctgct ggaccgcctg
     5221 gaggactggc tgccggacgt ggacacgctg ctggccggcc gcgaaccccg gttcgtccac
     5281 ggcgacctgc acgggaccaa catcttcgtg gacctggccg cgaccgaggt caccgggatc
     5341 gtcgacttca ccgacgtcta tgcgggagac tcccgctaca gcctggtgca actgcatctc
     5401 aacgccttcc ggggcgaccg cgagatcctg gccgcgctgc tcgacggggc gcagtggaag
     5461 cggaccgagg acttcgcccg cgaactgctc gccttcacct tcctgcacga cttcgaggtg
     5521 ttcgaggaga ccccgctgga tctctccggc ttcaccgatc cggaggaact ggcgcagttc
     5581 ctctgggggc cgccggacac cgcccccggc g
//
