LOCUS       Exported                3391 bp ds-DNA     circular SYN 10-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3391)
  AUTHORS   Kaiyi Jiang
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 13, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3391
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=gibson5'
     primer_bind     1..25
                     /label=AATII
     primer_bind     complement(1..25)
                     /label=AATIIR
     primer_bind     24..67
                     /label=KJ409
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    44..68
                     /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    77..235
                     /label=ORF
     misc_feature    377..397
                     /label=21nt guide
     ncRNA           398..465
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     398..414
                     /label=KJ309
     primer_bind     complement(444..465)
                     /label=Primer 7
     primer_bind     complement(444..465)
                     /label=Kj616
     primer_bind     complement(445..485)
                     /label=KJ140
     primer_bind     complement(445..485)
                     /label=Primer 2
     primer_bind     complement(445..465)
                     /label=Primer 5
     primer_bind     complement(445..465)
                     /label=KJ305
     misc_feature    456
                     /label=C>U inactive mutant
                     /label=nonstandard type: Misc
                     /note="/created_by=Lukas"
     terminator      474..521
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(521..542)
                     /label=KJ310
     primer_bind     complement(523..543)
                     /label=30_half_31_commonreverse
     CDS             857..1672
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(930..949)
                     /label=Kan-R
     primer_bind     1789..1806
                     /label=L4440
     rep_origin      2380..2968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3280..3299
                     /label=KJ778
ORIGIN
        1 tacttcgcgt ttgccacctg acgtctaata cgactcacta tagggaattg tgagcggata
       61 acaattcccc ggatccactt ccaagacctg tggtaattgc ggtgtgaaga acaacaaact
      121 tggtggaaag gaaacgttta cttgtgagtg ttgcaattac aaaactcatc gagacgtcaa
      181 cggagcgaga aacattctgt gcaaatactt gaaacttttt ccattcgcag cataacgaaa
      241 gaaactgaca atcgattttt tcgggttcga ttctatccca cttgactcaa agagtcagag
      301 ggctcgaata cattttctgc acaggttttg ctaatgcaag atctggggca agaatgtgtt
      361 cgggtcaaat gagttagcag ccacctcctt gttattgggc cggcatggtc ccagcctcct
      421 cgctggcgcc ggctgggcaa catgcttcgg catggcgaat gggacgcggc cgcctagcat
      481 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt ggctagcgct gaggtctgcc
      541 tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa
      601 agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa
      661 cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa
      721 ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc tctgatgtta
      781 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
      841 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga ggccgcgatt
      901 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
      961 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1021 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1081 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1141 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     1201 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1261 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1321 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     1381 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     1441 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     1501 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     1561 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     1621 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     1681 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggact cgagattacc
     1741 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1801 agcgaggaag cctgcataac gcgaagtaat cttttcggtt ttaaagaaaa agggcagggt
     1861 ggtgacacct tgcccgtttt tttgccggac tgcagcaatg gcaacaacgt tgcgcaaact
     1921 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1981 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2041 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2101 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2161 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2221 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2281 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2521 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2581 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2641 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2701 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2761 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2821 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2881 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2941 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3001 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3061 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3121 cagcgaggtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     3181 cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcatcga tgcataatgt
     3241 gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccct cgagccgtca
     3301 attgtctgat tcgttaccaa ttatactagc tccggcaaaa aaacgggcaa ggtgtcacca
     3361 ccctgccctt tttctttaaa accgaaaaga t
//
