LOCUS       sequence-11468-e        6792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6792)
  AUTHORS   Steen EJ, Kang Y, Bokinsky G, Hu Z, Schirmer A, McClure A, Del 
            Cardayre SB, Keasling JD.
  TITLE     Microbial production of fatty-acid-derived fuels and chemicals from 
            plant biomass.
  JOURNAL   Nature. 2010 Jan 28;463(7280):559-62.
  PUBMED    20111002
REFERENCE   2  (bases 1 to 6792)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature."; 
            date: "2010-01-28"; volume: "463(7280)"; pages: "559-62"
COMMENT     Sequence Label: pKS104
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..1131)
                     /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"
     primer_bind     1093..1112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1209)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1415..1434
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(1520..1525,1526..1543,1544..1550)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1558..1574
                     /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)."
     terminator      4730..4801
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4817..4844
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5002..5590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5082..5101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5678..5772)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5802..6593,6594..6662))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6425..6444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6663..6767)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg
     1561 tgagcggata acaatttcag aattcaaaag atcttttaag aaggagatat acatatgaag
     1621 aaggtttggc ttaaccgtta tcccgcggac gttccgacgg agatcaaccc tgaccgttat
     1681 caatctctgg tagatatgtt tgagcagtcg gtcgcgcgct acgccgatca acctgcgttt
     1741 gtgaatatgg gggaggtaat gaccttccgc aagctggaag aacgcagtcg cgcgtttgcc
     1801 gcttatttgc aacaagggtt ggggctgaag aaaggcgatc gcgttgcgtt gatgatgcct
     1861 aatttattgc aatatccggt ggcgctgttt ggcattttgc gtgccgggat gatcgtcgta
     1921 aacgttaacc cgttgtatac cccgcgtgag cttgagcatc agcttaacga tagcggcgca
     1981 tcggcgattg ttatcgtgtc taactttgct cacacactgg aaaaagtggt tgataaaacc
     2041 gccgttcagc acgtaattct gacccgtatg ggcgatcagc tatctacggc aaaaggcacg
     2101 gtagtcaatt tcgttgttaa atacatcaag cgtttggtgc cgaaatacca tctgccagat
     2161 gccatttcat ttcgtagcgc actgcataac ggctaccgga tgcagtacgt caaacccgaa
     2221 ctggtgccgg aagatttagc ttttctgcaa tacaccggcg gcaccactgg tgtggcgaaa
     2281 ggcgcgatgc tgactcaccg caatatgctg gcgaacctgg aacaggttaa cgcgacctat
     2341 ggtccgctgt tgcatccggg caaagagctg gtggtgacgg cgctgccgct gtatcacatt
     2401 tttgccctga ccattaactg cctgctgttt atcgaactgg gtgggcagaa cctgcttatc
     2461 actaacccgc gcgatattcc agggttggta aaagagttag cgaaatatcc gtttaccgct
     2521 atcacgggcg ttaacacctt gttcaatgcg ttgctgaaca ataaagagtt ccagcagctg
     2581 gatttctcca gtctgcatct ttccgcaggc ggtgggatgc cagtgcagca agtggtggca
     2641 gagcgttggg tgaaactgac cggacagtat ctgctggaag gctatggcct taccgagtgt
     2701 gcgccgctgg tcagcgttaa cccatatgat attgattatc atagtggtag catcggtttg
     2761 ccggtgccgt cgacggaagc caaactggtg gatgatgatg ataatgaagt accaccaggt
     2821 caaccgggtg agctttgtgt caaaggaccg caggtgatgc tgggttactg gcagcgtccc
     2881 gatgctaccg atgaaatcat caaaaatggc tggttacaca ccggcgacat cgcggtaatg
     2941 gatgaagaag gattcctgcg cattgtcgat cgtaaaaaag acatgattct ggtttccggt
     3001 tttaacgtct atcccaacga gattgaagat gtcgtcatgc agcatcctgg cgtacaggaa
     3061 gtcgcggctg ttggcgtacc ttccggctcc agtggtgaag cggtgaaaat cttcgtagtg
     3121 aaaaaagatc catcgcttac cgaagagtca ctggtgactt tttgccgccg tcagctcacg
     3181 ggatacaaag taccgaagct ggtggagttt cgtgatgagt taccgaaatc taacgtcgga
     3241 aaaattttgc gacgagaatt acgtgacgaa gcgcgcggca aagtggacaa taaagcctga
     3301 actagtaagg aggaaacaga atgcgcccat tacatccgat tgattttata ttcctgtcac
     3361 tagaaaaaag acaacagcct atgcatgtag gtggtttatt tttgtttcag attcctgata
     3421 acgccccaga cacctttatt caagatctgg tgaatgatat ccggatatca aaatcaatcc
     3481 ctgttccacc attcaacaat aaactgaatg ggcttttttg ggatgaagat gaagagtttg
     3541 atttagatca tcattttcgt catattgcac tgcctcatcc tggtcgtatt cgtgaattgc
     3601 ttatttatat ttcacaagag cacagtacgc tgctagatcg ggcaaagccc ttgtggacct
     3661 gcaatattat tgaaggaatt gaaggcaatc gttttgccat gtacttcaaa attcaccatg
     3721 cgatggtcga tggcgttgct ggtatgcggt taattgaaaa atcactctcc catgatgtaa
     3781 cagaaaaaag tatcgtgcca ccttggtgtg ttgagggaaa acgtgcaaag cgcttaagag
     3841 aacctaaaac aggtaaaatt aagaaaatca tgtctggtat taagagtcag cttcaggcga
     3901 cacccacagt cattcaagag ctttctcaga cagtatttaa agatattgga cgtaatcctg
     3961 atcatgtttc aagctttcag gcgccttgtt ctattttgaa tcagcgtgtg agctcatcgc
     4021 gacgttttgc agcacagtct tttgacctag atcgttttcg taatattgcc aaatcgttga
     4081 atgtgaccat taatgatgtt gtactagcgg tatgttctgg tgcattacgt gcgtatttga
     4141 tgagtcataa tagtttgcct tcaaaaccat taattgccat ggttccagcc tctattcgca
     4201 atgacgattc agatgtcagc aaccgtatta cgatgattct ggcaaatttg gcaacccaca
     4261 aagatgatcc tttacaacgt cttgaaatta tccgccgtag tgttcaaaac tcaaagcaac
     4321 gcttcaaacg tatgaccagc gatcagattc taaattatag tgctgtcgta tatggccctg
     4381 caggactcaa cataatttct ggcatgatgc caaaacgcca agccttcaat ctggttattt
     4441 ccaatgtgcc tggcccaaga gagccacttt actggaatgg tgccaaactt gatgcactct
     4501 acccagcttc aattgtatta gacggtcaag cattgaatat tacaatgacc agttatttag
     4561 ataaacttga agttggtttg attgcatgcc gtaatgcatt gccaagaatg cagaatttac
     4621 tgacacattt agaagaagaa attcaactat ttgaaggcgt aattgcaaag caggaagata
     4681 ttaaaacagc caattaagga tccaaactcg agtaaggatc tccaggcatc aaataaaacg
     4741 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4801 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata cctagggcgt
     4861 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4921 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4981 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5041 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5101 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5161 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5221 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5281 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5341 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5401 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5461 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5521 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5581 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5641 ctcacgttaa gggattttgg tcatgactag tgcttggatt ctcaccaata aaaaacgccc
     5701 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     5761 tcaacaggag tccaagcgag ctcgtaaact tggtctgaca gttaccaatg cttaatcagt
     5821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6781 aaagtgccac ct
//
