LOCUS       Exported                5667 bp ds-DNA     circular SYN 19-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  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..5667
                     /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..2110
                     /note="YTH domain of YTHDC2"
     CDS             1658..1675
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      2174..2221
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2258..2713
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2806..3621)
                     /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      3743..4331
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4517..4659
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4761..4952)
                     /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 aagcggtaac atgcctgttc gatacttcat aatgaagagt agcaatttga gaaaccttga
     1741 aatttctcaa cagaagggta tctggtctac aactcctagt aatgaacgga agctaaatcg
     1801 agccttttgg gaaagcagca tagtttactt ggtattttct gttcaaggat ctggacattt
     1861 ccagggattt tctaggatgt cttctgagat tggaagggaa aagagtcagg actggggctc
     1921 tgctggacta ggaggagtat ttaaggtgga gtggatacga aaagaaagcc ttccctttca
     1981 atttgcacac catttactca atccatggaa tgacaacaag aaagtgcaga taagcaggga
     2041 tgggcaggaa ctagaacctc tggttggtga acagttgctc cagttatggg aacgtcttcc
     2101 cttgggataa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2161 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2221 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     2281 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2341 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2401 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2461 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2521 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2581 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2701 acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2761 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     2821 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2881 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2941 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     3001 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     3061 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     3121 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     3181 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3241 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3301 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3361 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3421 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3481 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3541 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     3601 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     3661 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3721 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3781 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3841 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4201 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4261 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4321 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4381 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4441 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4501 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4561 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4621 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4681 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4741 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4801 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4861 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4921 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4981 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     5041 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     5101 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     5161 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     5221 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     5281 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     5341 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     5401 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     5461 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     5521 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     5581 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     5641 gcgacgatag tcatgccccg cgcccac
//
