LOCUS       Exported                6611 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTrcAgBIS (CO)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  AUTHORS   Peralta-Yahya PP, Ouellet M, Chan R, Mukhopadhyay A, Keasling JD, 
            Lee TS
  TITLE     Identification and microbial production of a terpene-based advanced 
            biofuel.
  JOURNAL   Nat Commun. 2011 Sep 27;2:483. doi: 10.1038/ncomms1494.
  PUBMED    21952217
REFERENCE   2  (bases 1 to 6611)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6611
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(2794..2811)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2964..3050
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3142..3169
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3189..3280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3281..4141
                     /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"
     primer_bind     complement(3499..3518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4312..4900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4801..4820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5054..5071
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5086..5226
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5152..5171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5312..5334
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5412..5489
                     /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             5490..6572
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5509..5528)
                     /label=LacI-R
                     /note="LacI, reverse primer"
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 gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg cgggtgtttc tgcggtttct aaagtttctt
      301 ctctggtttg cgacctgtct tctacctctg gtctgatccg tcgtaccgcg aacccgcacc
      361 cgaacgtttg gggttacgac ctggttcact ctctgaaatc tccgtacatc gactcttctt
      421 accgtgaacg tgcggaagtt ctggtttctg aaatcaaagc gatgctgaac ccggcgatca
      481 ccggtgacgg tgaatctatg atcaccccgt ctgcgtacga caccgcgtgg gttgcgcgtg
      541 ttccggcgat cgacggttct gcgcgtccgc agttcccgca gaccgttgac tggattctga
      601 aaaaccagct gaaagacggt tcttggggta tccagtctca cttcctgctg tctgaccgtc
      661 tgctggcgac cctgtcttgc gttctggttc tgctgaaatg gaacgttggt gacctgcagg
      721 ttgaacaggg tatcgagttc atcaaatcta acctggaact ggttaaagac gaaaccgacc
      781 aggactctct ggttaccgac ttcgaaatca tcttcccgtc tctgctgcgt gaagcgcagt
      841 ctctgcgtct gggtctgccg tacgacctgc cgtacatcca cctgctgcag accaaacgtc
      901 aggaacgtct ggcgaaactg tctcgtgaag aaatctacgc ggttccgtct ccgctgctgt
      961 actctctgga aggtatccag gacatcgttg aatgggaacg tatcatggaa gttcagtctc
     1021 aggacggttc tttcctgtct tctccggcgt ctaccgcgtg cgttttcatg cacaccggtg
     1081 acgcgaaatg cctggagttc ctgaactctg ttatgatcaa attcggtaac ttcgttccgt
     1141 gcctgtaccc ggttgacctg ctggaacgtc tgctgatcgt tgacaacatc gttcgtctgg
     1201 gtatctaccg tcacttcgaa aaagaaatca aagaagcgct ggactacgtt taccgtcact
     1261 ggaacgaacg tggtatcggt tggggtcgtc tgaacccgat cgcggacctg gaaaccaccg
     1321 cgctgggttt ccgtctgctg cgtctgcacc gttacaacgt ttctccggcg atcttcgaca
     1381 acttcaaaga cgcgaacggt aaattcatct gctctaccgg tcagttcaac aaagacgttg
     1441 cgtctatgct gaacctgtac cgtgcgtctc agctggcgtt tccgggtgaa aacatcctgg
     1501 acgaagcgaa atctttcgcg accaaatacc tgcgtgaagc gctggaaaaa tctgaaacct
     1561 cttctgcgtg gaacaacaaa cagaacctgt ctcaggaaat caaatacgcg ctgaaaacct
     1621 cttggcacgc gtctgttccg cgtgttgaag cgaaacgtta ctgccaggtt taccgtccgg
     1681 actacgcgcg tatcgcgaaa tgcgtttaca aactgccgta cgttaacaac gaaaaattcc
     1741 tggaactggg taaactggac ttcaacatca tccagtctat ccaccaggaa gaaatgaaaa
     1801 acgttacctc ttggttccgt gactctggtc tgccgctgtt caccttcgcg cgtgaacgtc
     1861 cgctggagtt ctacttcctg gttgcggcgg gtacctacga accgcagtac gcgaaatgcc
     1921 gtttcctgtt caccaaagtt gcgtgcctgc agaccgttct ggacgacatg tacgacacct
     1981 acggtaccct ggacgaactg aaactgttca ccgaagcggt tcgtcgttgg gacctgtctt
     2041 tcaccgaaaa cctgccggac tacatgaaac tgtgctacca gatttactac gacatcgttc
     2101 acgaagttgc gtgggaagcg gaaaaagaac agggtcgtga actggtttct ttcttccgta
     2161 aaggttggga agactacctg ctgggttact acgaagaagc ggaatggctg gcggcggaat
     2221 acgttccgac cctggacgaa tacatcaaaa acggtatcac ctctatcggt cagcgtatcc
     2281 tgctgctgtc tggtgttctg atcatggacg gtcagctgct gtctcaggaa gcgctggaaa
     2341 aagttgacta tccgggtcgt cgtgttctga ccgaactgaa ctctctgatc tctcgtctgg
     2401 cggacgacac caaaacctac aaagcggaaa aagcgcgtgg tgaactggcg tcttctatcg
     2461 aatgctacat gaaagaccac ccggaatgca ccgaagaaga agcgctggac cacatctact
     2521 ctatcctgga accggcggtt aaagaactga cccgtgagtt cctgaaaccg gacgacgttc
     2581 cgttcgcgtg caaaaaaatg ctgttcgaag aaacccgtgt tactatggtt atcttcaaag
     2641 acggtgacgg tttcggtgtt tctaaactgg aagttaaaga ccacatcaaa gaatgcctga
     2701 tcgaaccgct gccgctgtaa cccggggatc ctctagagtc gacctgcagg catgcaagct
     2761 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     2821 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     2881 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     2941 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     3001 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     3061 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     3121 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     3181 aaactctttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3241 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3301 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3361 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3421 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3481 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     3541 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3601 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3661 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3721 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3781 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctacagcaa tggcaacaac
     3841 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3901 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3961 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4021 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4081 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4141 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4201 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4441 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4921 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4981 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5041 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     5101 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     5161 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     5221 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5281 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5341 ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat
     5401 gcatttacgt tgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
     5461 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     5521 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     5581 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     5641 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     5701 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     5761 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     5821 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     5881 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     5941 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     6001 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     6061 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     6121 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     6181 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     6241 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     6301 tgggatacga cgataccgaa gacagctcat gttatatccc gccgtcaacc accatcaaac
     6361 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     6421 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
     6481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagcgc
     6601 gaattgatct g
//
