LOCUS       Exported                5832 bp ds-DNA     circular SYN 19-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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..5832
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(94..1176)
                     /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"
     promoter        complement(1177..1254)
                     /gene="lacI"
                     /note="lacI promoter"
     promoter        1567..1585
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1586..1610
                     /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)."
     RBS             1625..1647
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    1655..2275
                     /note="YTH domain of YTHDF3"
     CDS             1658..1675
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      2339..2386
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2423..2878
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2971..3786)
                     /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      3908..4496
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4682..4824
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4926..5117)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
       61 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
      121 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
      181 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
      241 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
      301 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
      361 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
      421 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
      481 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
      541 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
      601 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
      661 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
      721 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
      781 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
      841 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
      901 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
      961 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     1021 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     1081 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     1141 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     1201 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     1261 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     1321 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     1381 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     1441 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     1501 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     1561 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     1621 taattttgtt taactttaag aaggagatat acatatgcac catcatcatc atcatggtgg
     1681 aagcggtagt gtagaagtgc atcccgtgct ggaaaagcta aaggccataa acaactataa
     1741 tcccaaagac tttgattgga atctgaagaa tggacgtgtg tttataatta aaagctactc
     1801 tgaggatgac atacatcgtt ccattaaata ctctatctgg tgtagtactg agcatggtaa
     1861 taagcgtttg gatgcagctt accgttccct gaatgggaaa ggcccactct atttactctt
     1921 cagtgtgaat ggcagtggac atttttgtgg agtggctgaa atgaagtctg ttgtggacta
     1981 taatgcgtat gctggtgtct ggtctcagga taagtggaag ggcaaatttg aagttaaatg
     2041 gatctttgtc aaagatgttc ccaataacca attacggcat attcgcttag aaaataatga
     2101 caacaaaccg gttaccaatt caagggacac tcaagaggta cccctagaaa aagctaagca
     2161 agtgcttaaa ataattgcta ctttcaagca taccacctca atctttgatg actttgcaca
     2221 ttatgaaaag cgtcaagaag aggaggaagc catgcgtagg gagagaaata gataagatcc
     2281 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2341 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     2401 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2461 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2521 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2581 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2641 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2701 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2761 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2821 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttca
     2881 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2941 tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa
     3001 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     3061 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     3121 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     3181 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     3241 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     3301 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     3361 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     3421 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     3481 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     3541 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     3601 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     3661 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     3721 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     3781 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca
     3841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4021 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4621 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4681 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg
     4741 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4801 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4861 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4921 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     4981 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     5041 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     5101 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     5161 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     5221 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     5281 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     5341 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     5401 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     5461 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     5521 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     5581 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     5641 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     5701 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     5761 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     5821 ccccgcgccc ac
//
