LOCUS       190821_pET24a-nc        8336 bp DNA     circular SYN 18-OCT-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    ncOGT-FL_codon optimized for E.Coli_pET24a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8336)
  AUTHORS   Yi Zhu
  TITLE     Direct Submission
  JOURNAL   Exported Oct 18, 2022 from SnapGene Viewer 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(560..1375)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1497..2085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    2271..2413
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(2515..2706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3515..4597)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4598..4675)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    5003..5027
                     /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)."
                     /note="color: #31849b"
     RBS             5042..5064
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             5075..5092
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             5093..8200
                     /label=Full-length human ncOGT_codon optimized for E.coli
                     /note="color: #00ccff; direction: RIGHT"
     terminator      8264..8311
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatataca tatgcaccac catcatcatc atgcaagcag
     5101 cgttggtaat gttgcagata gcaccggtct ggcagaactg gcacatcgtg aatatcaggc
     5161 aggcgatttt gaagcagcag aacgtcattg tatgcagctg tggcgtcagg aaccggataa
     5221 tacaggcgtg ctgctgctgc tgagcagcat tcattttcag tgtcgtcgtc tggatcgtag
     5281 cgcacatttt agcaccctgg caattaaaca gaatccgctg ctggcagaag catatagcaa
     5341 tctgggtaat gtgtataaag aacgtggtca gctgcaggaa gcaattgaac attatcgtca
     5401 tgcactgcgt ctgaaaccgg attttattga tggctatatt aatctggcag cagcactggt
     5461 tgcagccggt gatatggaag gtgcagttca ggcatatgtt agcgcactgc agtataatcc
     5521 ggatctgtat tgtgttcgta gtgatctggg taatctgctg aaagcactgg gtcgtctgga
     5581 agaagcaaaa gcctgttatc tgaaagcaat tgaaacccag ccgaattttg cagttgcatg
     5641 gtcaaatctg ggttgcgttt ttaatgcaca gggtgaaatt tggctggcca ttcatcattt
     5701 tgaaaaagca gttaccctgg atccgaattt tctggatgca tatattaatc tgggcaatgt
     5761 gctgaaagaa gcccgtattt ttgatcgtgc agttgcagca tatctgcgtg cactgagcct
     5821 gagcccgaat catgcagttg ttcatggtaa tctggcctgc gtttattatg aacagggtct
     5881 gattgatctg gccattgata cctatcgtcg tgcaattgaa ctgcagccgc attttccgga
     5941 tgcctattgc aatctggcaa atgcactgaa agaaaaaggt agcgttgcag aagcagaaga
     6001 ttgttataat accgcactgc gcctgtgtcc gacccatgca gatagcctga ataatctggc
     6061 caatattaaa cgcgaacagg gcaatattga agaagcagtt cgtctgtatc gtaaagccct
     6121 ggaagttttt ccggaatttg cagcagccca tagtaatctg gcaagcgttc tgcagcagca
     6181 gggtaaactg caggaagccc tgatgcatta taaagaagca attcgtatta gcccgacctt
     6241 tgcagatgcc tatagtaata tgggtaatac cctgaaagaa atgcaggatg ttcagggtgc
     6301 actgcagtgt tatacccgtg caattcagat taatccggca tttgcagatg cacattcaaa
     6361 tctggccagc attcataaag atagcggtaa tattccggaa gccattgcaa gctatcgtac
     6421 cgcactgaaa ctgaaaccgg attttccgga cgcatattgt aatctggccc attgtctgca
     6481 gattgtttgt gattggaccg attatgatga acgcatgaaa aaactggtta gcattgttgc
     6541 agatcagctg gaaaaaaatc gtctgccgag cgttcatccg catcatagca tgctgtatcc
     6601 gctgagccat ggttttcgta aagcaattgc cgaacgtcat ggcaatctgt gtctggataa
     6661 aattaatgtg ctgcataaac cgccgtatga acatccgaaa gatctgaaac tgagtgatgg
     6721 tcgtctgcgt gttggttatg ttagcagcga ttttggtaat catccgacca gccatctgat
     6781 gcagagcatt ccgggtatgc ataatccgga taaatttgaa gttttttgct atgccctgag
     6841 tccggatgat ggcaccaatt ttcgtgttaa agttatggca gaagccaatc attttattga
     6901 tctgagccag attccgtgca atggtaaagc agcagatcgt attcatcagg atggtattca
     6961 tattctggtg aatatgaatg gctataccaa aggtgcacgc aatgaactgt ttgcactgcg
     7021 tccggcaccg attcaggcaa tgtggctggg ttatccgggt acaagcggtg cactgtttat
     7081 ggattatatt attaccgatc aggaaaccag tccggcagaa gttgcagaac agtatagcga
     7141 aaaactggcc tatatgccgc ataccttttt tattggtgat catgccaata tgtttccgca
     7201 tctgaaaaaa aaagccgtga ttgattttaa aagcaatggc catatttatg ataatcgcat
     7261 tgtgctgaat ggcattgatc tgaaagcatt tctggatagc ctgccggatg ttaaaattgt
     7321 gaaaatgaaa tgtccggatg gtggcgataa tgcagatagc agcaatacag cactgaatat
     7381 gccggttatt ccgatgaata ccattgccga agcagtgatt gaaatgatta atcgtggtca
     7441 gattcagatt accattaatg gctttagcat tagcaatggt ctggcaacca cacagattaa
     7501 taataaagca gcaaccggtg aagaagttcc gcgtaccatt attgttacca cccgtagcca
     7561 gtatggtctg ccggaagatg caattgttta ttgcaatttt aatcagctgt ataaaattga
     7621 tccgagcacc ctgcagatgt gggcaaatat tctgaaacgt gttccgaata gcgttctgtg
     7681 gctgctgcgt tttccggcag ttggtgaacc gaatattcag cagtatgcac agaatatggg
     7741 tctgccgcag aatcgcatta tttttagtcc ggttgcaccg aaagaagaac atgttcgtcg
     7801 cggtcagctg gcagatgttt gtctggatac accgctgtgt aatggtcata ccaccggtat
     7861 ggatgttctg tgggcaggca ccccgatggt taccatgccg ggtgaaaccc tggcaagccg
     7921 tgttgcagca agccagctga cctgtctggg ttgtctggaa ctgattgcaa aaaatcgtca
     7981 ggaatatgaa gatattgccg ttaaactggg caccgatctg gaatatctga aaaaagttcg
     8041 tggcaaagtt tggaaacagc gtattagcag tccgctgttt aataccaaac agtataccat
     8101 ggaactggaa cgtctgtatc tgcagatgtg ggaacattat gcagcaggta ataaaccgga
     8161 tcatatgatt aaaccggttg aagttaccga aagcgcctaa gatccggctg ctaacaaagc
     8221 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     8281 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggat
//
