LOCUS       Exported                5296 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 5296)
  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..5296
                     /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..863
                     /codon_start=1
                     /label=RHEB GTPase
                     /translation="MPQSKSRKIAILGYCSVGKSSLTIQFVEGQFVDSYDPTIENTFTK
                     LITVNGQEYHLQLVDTAGQDEYSIFPQTYSIDINGYILVYSVTSIKSFEVIKVIHGKLL
                     DMVGKVQIPIMLVGNKKDLHMERVISYEEGKALAESWNAAFLESSAKENQTAVDVFRRI
                     ILEAEKM"
     terminator      1155..1241
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1333..1360
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1380..1471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1472..2332
                     /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      2503..3091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(3339..3767)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4097..4174
                     /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             4175..5257
                     /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 atgccccaat
      361 caaaaagtag gaagatagct atcctcggtt actgcagcgt aggcaagtct agcttgacca
      421 ttcagttcgt ggaaggtcaa ttcgtggaca gctatgatcc gacgatcgag aacaccttca
      481 ccaaactgat taccgttaat ggccaagaat accacctgca gctggtggac accgctggtc
      541 aagatgagta ttcgatcttt ccgcagacgt actctatcga catcaacggc tacattttgg
      601 tgtattccgt gaccagcatc aagagctttg aagttatcaa agtcattcat ggtaagctgc
      661 tggatatggt tggtaaggtg cagattccga ttatgctggt cgggaacaaa aaagatctgc
      721 acatggaacg tgttatcagc tatgaagagg gcaaggcgct ggcagagtcc tggaatgcgg
      781 cgtttttgga gagctccgcg aaagaaaacc agactgccgt tgacgttttc cgccgtatta
      841 tccttgaggc agagaaaatg taagaattca aaggcctacg tcgacgagct cactagtcgc
      901 ggccgctttc gaatctagag cctgcagtct cgaggcatgc ggtaccaagc ttggctgttt
      961 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     1021 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     1081 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     1141 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1201 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     1261 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     1321 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     1381 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1441 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1501 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1561 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1621 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1681 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     1741 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1801 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1861 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1921 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1981 cataccaaac gacgagcgtg acaccacgat gcctacagca atggcaacaa cgttgcgcaa
     2041 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2101 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2161 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2221 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2281 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2341 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2401 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2461 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2521 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2581 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2641 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2701 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2761 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2821 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2881 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2941 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3001 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3061 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3121 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3181 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3241 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3301 gcatctgtgc ggtatttcac accgcataat tttgttaaaa ttcgcgttaa atttttgtta
     3361 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3421 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3481 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3541 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3601 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3661 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3721 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     3781 caggctgcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3841 ccagtcacgt agcgatatcg ctacgtgact ggtacactcc gctatcgcta cgtgactggg
     3901 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3961 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4021 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc
     4081 ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata
     4141 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     4201 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     4261 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     4321 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     4381 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     4441 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     4501 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     4561 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     4621 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     4681 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4741 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4801 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4861 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     4921 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     4981 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     5041 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     5101 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     5161 cctggcaccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5221 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     5281 agcgcgaatt gatctg
//
