LOCUS       Exported                6326 bp DNA     circular SYN 22-JUL-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6326
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        471..575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             576..1436
                     /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"
     rep_origin      1607..2195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2483..2504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2519..2549
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2557..2573
                     /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        2618..2636
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2663..3662
                     /label=H.S. PABPC1 5' homology arm
     sig_peptide     3696..4376
                     /product=" missing first two METs and stop codon"
                     /label=A.T. IAA17/AXR3 (AID)
     CDS             3696..4376
                     /codon_start=1
                     /gene="Arabidopsis thaliana AXR3"
                     /product="auxin-responsive IAA17 protein"
                     /label=auxin-inducible degron
                     /note="acts as an auxin-inducible degron (AID) in the
                     presence of TIR1"
                     /translation="GSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNNE
                     PANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGG
                     PEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMNE
                     RKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAMEK
                     CKSRA"
     misc_feature    4377..4430
                     /label=T2A
     misc_feature    4443..5153
                     /label=mCherry
                     /note="note=""direct strand 
                     translation""translation=""MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEF
                      
                     EIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLS
                      
                     FPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEA
                      
                     SSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSH
                      "
     misc_feature    5154..6108
                     /label=H.S. PABPC1 3' homology arm
     misc_feature    5322..5341
                     /label=CRISPR guide RNA target site-1
     misc_feature    5342..5344
                     /label=Mutated PAM-1
     promoter        complement(6133..6151)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6158..6174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(join(6315..6326,1..444))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
       61 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1081 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1261 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2341 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2401 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2461 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     2521 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     2581 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
     2641 aaaagctggt accgggcccc ccgaacaccg atttaacaac ctaatggtaa cattgcttaa
     2701 ttggcaagct gataatatca agagttaatt gccttgtggc agattttatt ctgtcttccc
     2761 atgaagacat aatgaaattg tggattatgg gttattatat acgtgagaca gaagtatggt
     2821 tcttaaaccc cagtttaatg tggtaattca atgcagttaa aaactttttt ttattgaagg
     2881 gagtttttga atgtttctgt tcctaggtga acggctgttt cctcttattc aagccatgca
     2941 ccctactctt gctggtaaaa tcactggcat gttgttggag attgataatt cagaacttct
     3001 tcatatgctc gagtctccag agtcactccg ttctaaggtg atttaaattt atttaccaag
     3061 ttgcctctta cacagttatt acgtggcttt ttatggggac ttaattgtac agaacactta
     3121 catcgtgtta tatataagcc atgtacagtt gtaaagtctg cataataacc tgtggcgcta
     3181 tttatcccca ttttatagat aggtaaactg aaacaggctg aataatttgc caaatgctca
     3241 cacagctctt aagtggcaga gcctggcttc aaatcccatc agctccaaca gccatactct
     3301 caactgctgc tgctttctct agtggagaaa gagaaatatt tttcgagtgt ggggaggaca
     3361 aggaaagagt agtgcagggg agcgattcat aattctagaa tagtggtgta atagaactca
     3421 aaattgaatg gatatagttg tttagtgtaa caatttatac tttcatttga gggaagaaac
     3481 ttatctttgt tgggaattgg tatgttacca ttatgaaact ctttatttcc attcctttcc
     3541 tgaaccagtt tatctgtttt cccctttttt aggttgatga agctgtagct gtactacaag
     3601 cccaccaagc taaagaggct gcccagaaag cagttaacag tgccaccggt gttccaactg
     3661 ttgtcgactc cggagccgcc gctgctggcg gatctggcag tgtcgagctg aatctgaggg
     3721 agactgagct gtgtcttggt cttcccggtg gagatacagt ggctccggta accggaaaca
     3781 agagagggtt ctcagagacg gttgatctga agctaaatct gaataatgag cctgcaaaca
     3841 aggaaggatc tacgactcat gacgtcgtga cttttgattc caaggagaag agtgcttgtc
     3901 ctaaagatcc agccaaacct ccggccaagg cacaagttgt gggatggcca ccggtgagat
     3961 cataccggaa gaacgtgatg gtttcctgcc aaaaatcaag cggtggcccg gaggcggcgg
     4021 cgttcgtgaa ggtatcaatg gacggagcac cgtacttgag gaaaatcgat ttgaggatgt
     4081 ataaaagcta cgatgagctt tctaatgctt tgtccaacat gttcagctct tttaccatgg
     4141 gcaaacatgg aggagaagaa ggaatgatag acttcatgaa tgagaggaaa ttgatggatt
     4201 tggtgaatag ctgggactat gttccctctt atgaagacaa agacggtgat tggatgctcg
     4261 tcggcgacgt tccttggcca atgttcgtcg atacatgcaa gcgtttacgt ctcatgaaag
     4321 gatcggatgc cattggtctc gctccgaggg cgatggagaa gtgcaagagc agagctgagg
     4381 gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca gcggccgcaa
     4441 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     4501 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     4561 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     4621 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     4681 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     4741 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tctctccagg
     4801 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     4861 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     4921 gcgctctgaa aggagagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     4981 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     5041 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     5101 aacgcgccga gggacggcac tccaccggag gaatggacga gctgtacaaa taaagtgagc
     5161 ttttccctct ggtttgttgt tttaagcttg ttttcttcat cttttataaa gtttgttgag
     5221 catattttga aatgtacata gcaaaagatc attttacagt tcatttaaat gtattgctaa
     5281 tttttcaagg aaaagtggaa ctaactatag aggaattaaa ggaattttac ccgttaactg
     5341 ttttattttt aaaatcccag gactttcagg tggtgttttt tttttttttt ttttttgaga
     5401 tggagtttcg ttctgtcgcc caacctggag tgcattgacg cgatctcggc tcactgcaac
     5461 ctccacctcc caggttcaag caattctcct gcctcggcct cccgagcagc tgggattaac
     5521 aggcccatgc ctgccaccgt gcccagctaa tttttgtatt tttactagac ttggggtttt
     5581 gccatgttgg ccaggctggt ctggaacttc tgacctcaag tgatccgccc acctcggcct
     5641 cccaaagtct tgaaattaca ggcgtgagcc actgcgccct gccagatttt gtatagtaat
     5701 atatatataa aatgacctgt cacaggttgg ctttcatagt tttacctttc ctttttgggt
     5761 ggtgcccttg ggtctctact actgtctttc atcactccca agaaattgag tagtgcagcc
     5821 aggatcagga ccagggattg gcaaatgttt tttgtaaagg gccagatagg aaatattttg
     5881 ggctttgagg actgtgttat ctctgaaatg atcttgttcc agtgttacca tgtcaagaca
     5941 accatagata gtatgtatac aaatgaacag gtatggctgt gttccagtaa aactatggat
     6001 aaaaacaacc acattgcatt tggctaaggg ctttaatttg ctactgacaa gtgatagtcc
     6061 tggtcttaat ttgttttttt aacatttaca gattgatcag ggaccatgcg ccgcggtgga
     6121 gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     6181 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6241 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6301 tgaatggcga atggaaattg taagcg
//
