LOCUS       Exported                5284 bp ds-DNA     circular SYN 15-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5284)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5284
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    230..246
                     /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)."
     CDS             279..296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..338
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             351..848
                     /codon_start=1
                     /label=NRAS_CL_G12C
                     /translation="MTEYKLVVVGACGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETSLLDILDTAGQEEYSAMRDQYMRTGEGFLLVFAINNSKSFADINLYREQIKRVK
                     DSDDVPMVLVGNKSDLPTRTVDTKQAHELAKSYGIPFIETSAKTRQGVEDAFYTLVREI
                     RQF"
     terminator      1143..1229
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1321..1348
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1368..1459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1460..2320
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2491..3079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(3327..3755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4085..4162
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4163..5245
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gtccgtataa tctgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgt cgtactacca tcaccatcac catcacgatt
      301 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc atgacggaat
      361 ataagctggt ggtggtgggc gcctgcggtg tgggcaagag tgcgctgacc atccagctga
      421 tccagaacca ttttgtggac gaatacgacc ccactataga ggattcctac cggaagcagg
      481 tggtcattga tggggagacg agcctgttgg acatcctgga taccgccggc caggaggagt
      541 acagcgccat gcgggaccag tacatgcgca ccggggaggg cttcctgctg gtgtttgcca
      601 tcaacaactc taagtctttt gcggacatca acctgtacag ggagcagatc aaacgggtga
      661 aggactcgga tgacgtgccc atggtgctgg tggggaacaa gagtgacctg cctacacgca
      721 ctgtggatac taagcaggct cacgaactcg ccaaaagcta cggcatcccc ttcatcgaga
      781 cctcggccaa gacccggcag ggagtggagg atgccttcta cacgttggtg cgtgagatcc
      841 ggcagttctg agaattcaaa ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga
      901 atctagagcc tgcagtctcg aggcatgcgg taccaagctt ggctgttttg gcggatgaga
      961 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     1021 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     1081 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     1141 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1201 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     1261 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     1321 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttt gtttattttt
     1381 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1441 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1501 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1561 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1621 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1681 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     1741 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1801 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1861 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1921 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1981 cgagcgtgac accacgatgc ctacagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2041 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2101 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2161 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2221 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2281 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2341 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2401 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2461 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2521 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2581 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2641 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2701 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2761 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2821 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2881 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2941 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3001 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3061 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3121 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3181 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3241 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3301 tatttcacac cgcataattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3361 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3421 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3481 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3541 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3601 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3661 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3721 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgctatg
     3781 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtacc agtcacgtag
     3841 cgatatcgct acgtgactgg tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3901 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3961 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4021 caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta
     4081 cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     4141 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     4201 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     4261 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     4321 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     4381 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     4441 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     4501 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     4561 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     4621 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     4681 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     4741 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     4801 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     4861 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     4921 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     4981 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     5041 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     5101 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcacccaa
     5161 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5221 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag cgcgaattga
     5281 tctg
//
