LOCUS       Exported                5760 bp ds-DNA     circular SYN 19-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5760)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Oct 19, 2024 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5760
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      17..64
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    128..676
                     /note="YTH domain of YTHDF2"
     CDS             complement(656..673)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     RBS             684..706
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    721..745
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(746..764)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1077..1154
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             1155..2237
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             3046..3237
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3339..3481
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3667..4255)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4377..5192
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5285..5740)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
       61 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
      121 ccggatctta aatggaagtg gtgtgcttgt agctggctat aattttcaac acctgcttag
      181 ccttttccag aggcacttcc tgagtgtccc tagagttggt cactggttta ttctcgttgt
      241 tctctaggcg aatgtgtcgc agttggctat tgggaacgtc cttcacaaaa atccacctga
      301 catcaaaacg acccttccat ttgtcctggg accacacacc tgcacatgtg ttgtagtcca
      361 cagcagattt catttctgcc acgccacaga agtgtccact gccgttgaca ctgaaaagta
      421 agtaaacggg gcctttcccg ttcatggaac gataagcagc atccagtctc ttgttaccat
      481 gctctgtgct gcaccaaata ttatacttaa tggaacggtg aatatcgtcc tcagagtagc
      541 tcttaatgat gaaaacccgg ccatgtttca gattccagtc aaaatctttg gggttatagt
      601 tattaatgga ccgaagcttc tccaacactg ggtggggttc tgaaccgctt ccaccatgat
      661 gatgatgatg gtgcatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg
      721 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
      781 atcgatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
      841 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
      901 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
      961 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     1021 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca
     1081 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     1141 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     1201 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     1261 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     1321 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     1381 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     1441 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     1501 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     1561 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     1621 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     1681 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     1741 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     1801 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     1861 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     1921 ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga tacgacgata
     1981 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     2041 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     2101 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     2161 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2221 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
     2281 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     2341 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     2401 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     2461 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     2521 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca
     2581 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     2641 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     2701 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     2761 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     2821 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     2881 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     2941 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     3001 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     3061 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     3121 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     3181 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3241 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3301 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3361 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     3421 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4081 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4321 tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     4381 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
     4441 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     4501 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     4561 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     4621 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     4681 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     4741 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     4801 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     4861 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     4921 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     4981 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     5041 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     5101 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     5161 agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt
     5221 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5281 cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5341 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5401 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5461 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5521 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5581 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5641 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5701 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc aatccggata
//
