LOCUS       sequence-45433-e        8589 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 8589)
  AUTHORS   Bokinsky G, Peralta-Yahya PP, George A, Holmes BM, Steen EJ, 
            Dietrich J, Soon Lee T, Tullman-Ercek D, Voigt CA, Simmons BA, 
            Keasling JD
  TITLE     Synthesis of three advanced biofuels from ionic liquid-pretreated 
            switchgrass using engineered Escherichia coli.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Dec 13;108(50):19949-54. Epub 2011 
            Nov 28.
  PUBMED    22123987
REFERENCE   2  (bases 1 to 8589)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2011-12-13"; volume: "108(50)"; pages: 
            "19949-54. Epub 2011 Nov 28"
COMMENT     Sequence Label: pCellulose
FEATURES             Location/Qualifiers
     source          1..8589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(3131..3162)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      5118..5189
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5205..5232
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5763..6713)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(7475..7569)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(7599..8390,8391..8459))
                     /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     8222..8241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8460..8564)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg tctccaataa ttatccataa
       61 gccgctctta tgattaaaaa aacgcgatga gataaaaatg tgtcattttg cgacaaaatt
      121 acgtgcttgt gaaagttgtt ataaatcaaa taagtggttg tgaaatttgc actctgaaaa
      181 ggacgtctta tctttaaata agtggtagcg aatcgctacg gaatagagat aacacgagga
      241 gtggttagaa gaattcaaaa gatctaggag gaaaaaaaaa tgaaagatga tttccctcaa
      301 cgcttgttgc gcgtgagtgc cgcatcgctt ttggtgctgg cggcactggc gggttgtgat
      361 tcccgcgctc cctctacaac agactctacg acgacagtat ctgccgatac tgtgacgacc
      421 caggccaccg aaaaggtgga gtggccagtg ctcaacagcg ccattaaaaa agacccggca
      481 gtcgaggcgc gtgtggatga cttgctggcg cgcatgacac tggaagaaaa aatcggccaa
      541 ctggtccagc cggaaattcg ccatgtgacg cccgaggaca ttaagcaata ccacgtcggt
      601 tcggtcttaa acggcggtgg cagcacaccc ggtgccaaca aatatgccag ccttgaagac
      661 tgggtgaagc tggcagacag cttctattac gcctcggtgg ataaatctga tggtcgtatc
      721 ggtattccgg tgatctgggg taccgatgcg gtacacggcc tgggtaacgt tataggtgca
      781 accctgtttc cgcacaacat aggtctgggc gcgaccaaca acccggagtt gctcaagcaa
      841 atcggttggg cgacagcgcg tgaaattgct gcgactggcc tggattggga tttctcgcct
      901 acagtcgccg tagcgcgcga tgatcgctgg ggccgcacct acgaatcctg gtcggaagat
      961 ccgcagattg tccatgcctt cgccggcaag atggtggagg gactgcaggg cacaggaggt
     1021 tctgaccgac tgtttaccca tgaacatgtt atcgccacgg ccaagcactt tattggcgac
     1081 ggcggtacgt tgaatggtgt ggatcgcggt gaaacccagg gcgatgaaaa agtcctgcgc
     1141 gatatccacg gtgccggtta tttcagtgcg attgagtctg gcgtgcaggt ggtgatggcc
     1201 tcctttacca gttgggaagg cacgcgcatg cacggccaca agtacctgct gaccgatgtg
     1261 cttaaagacc gtatgggttt tgatggcctg gtggtggggg attggagcgg ccacagtttt
     1321 attcccggtt gtaccgcgct taattgccca caatcgctga tggcagggct ggatatttac
     1381 atggtgcccg agccggattg gaaagagctg tataaaaatc tgctggcgca ggccaaaacc
     1441 ggcgagcttc ccatggcgcg ggtcgacgac gcggtgcggc gcatcctgcg cgtgaaaatc
     1501 cgtgcgggcc tgtttgaaaa aggcgcacca tcgacgcgac cgctggctgg taaaaaagac
     1561 gtgctcggtg cccccgaaca ccgcgaggtt gcccgccagg ccgtgcgtga atccctggtg
     1621 ttgctgaaaa acaaaaataa tctcctgcct ctcgcccgcc agcaaaccgt gttggtcacc
     1681 ggggatggtg ccgacaatat cggcaagcaa tccggtggct ggtcagtgtc ctggcagggc
     1741 acgggcaaca ccaatgcgga cttccccggc gctacttcca tctatgccgg tatcaacgcc
     1801 gtggtggagc aggccggtgg taaaaccctg ctgagtgacg atggcagctt tagtgaaaaa
     1861 cccgatgtgg cgattgtggt attcggtgaa gatccctatg ccgaaatgca gggcgatgtg
     1921 ggcaacatgg cctataaacc acgcgacacc agcgattggg aactgttgaa gaaattacgc
     1981 agccagggta ttccggtggt atcgctgttt atttccggcc gcccactgtg ggtgaaccgc
     2041 gagatcaatg cctcggatgc gtttgtggcc gtctggttgc cgggcactga aggccagggg
     2101 attgccgatg tcatcttcag gaatgcgcag ggcgagatca attacgacgt gaaaggccgc
     2161 ctgagtttct cctggcccaa gcgccccgaa caaacaccgc tgaaccgcgg tgatgccaat
     2221 tacgatccgc tgttccccta tggctatggc ctgagctatg gcgataaaga taccctgggt
     2281 gacaacctgt ccgaggaggg tattcaattg gctgaggctc tggatgtgct cgacctgttc
     2341 aaccgtcgtc cgattgagcc ctggcaattg gaaatcatcg gttaccagaa tgaccgtgtg
     2401 cccatggcca gcagtacagt gaccgcttcc tcgctgaaaa ttcaggcggt ggatcgcaac
     2461 gagcaagagg acgcgcgccg cgtgcaatgg aatggctcgg gacctggcca ggttgccctg
     2521 tcagtgggtg atcgccagga ttttatcggc tacgttaaat ccgattcggc gctggtcttt
     2581 gatatcaagg tgaatgcggc tcccacagtg acgacttacc tgcgcttggg ttgcggttct
     2641 tactgcgcat cggatatcga cctgacggaa aaacttaaag gctttgccgg acaggattgg
     2701 cagacagtga ccgtgccact ccactgctac ccgaattccg gcgcaaactt cggtgtaacc
     2761 caaccgccgg aagagttctg gacccaggtg ctgcaaccct tctcactgtt aaccagtggc
     2821 acattggatg tgacctttgc gcaggtgcgt gtcgtaaaag gtgctggaaa agacgtcgcc
     2881 tgcccctaat aagtatggtc gacgatccgg caaacaaacc accgttggta gcggtggttt
     2941 ttttgtttgg atcgacaatc ttcgtaagcg tcatcaataa gcgtaaaaaa accgggcaat
     3001 gcccggtttt ttaatgagaa attttacctg tcgtagccgc caccatccgg caaagaagca
     3061 tacaaggctt ttggcttata gctacgtagc gcattgcgtc gcagcacaat cccggcaccg
     3121 atcgagtaat agtcaaaagc ctccggtcgg aggcttttga ctttctgccc ttgtcagtta
     3181 tcatcttcgg ttacggttat cggcagaacg ccctgccgct gacaagagct tatttgcctg
     3241 accaatctac tccacatgat agataactat catctatcgt tggcatcagc gacatctgtc
     3301 acattcctgt caatagcgtt aactgcttca aatttttgat tcatacctgc cccttgtccc
     3361 ctgtcaaatg cttgacggct cgccctaatt ctctaaattg tatttctaga gttggcgagg
     3421 ttttgaacag ccccctctct gaccccggtt tattccatct tacttgtata agatttgcga
     3481 aggatgtcga agcgaattca aaagatctag caagaggaaa aaaaaatgac tatgacaaga
     3541 ctgaagattt cgaaaactct gctggctgta atgttgacct ctgccgtcgc gaccggctct
     3601 gcctacgcgg aaaacaacgc gcagactacc aatgaaagcg cagggcaaaa agtcgatagc
     3661 tctatgaata aagtcggtaa tttcatggat gacagcgcca tcaccgcgaa agtgaaggcg
     3721 gccctggtgg atcatgacaa catcaagagc accgatatct ctgtaaaaac cgatcaaaaa
     3781 gtcgtgaccc tgagcggttt cgttgaaagc caggcccagg ccgaagaggc agtgaaagtg
     3841 gcgaaaggcg ttgaaggggt gacctctgtc agcgacaaac tgcacgttcg cgacgctaaa
     3901 gaaggctcgg tgaagggcta cgcgggtgac accgccacca ccagtgaaat caaagccaaa
     3961 ctgctggcgg acgatatcgt cccttcccgt catgtgaaag ttgaaaccac cgacggcgtg
     4021 gttcagctct ccggtaccgt cgattctcag gcacaaagtg accgtgctga aagtatcgcc
     4081 aaagcggtag atggtgtgaa aagcgttaaa aatgatctga aaactaaggg cagcggatct
     4141 atccgcagcc cagcgtccgc cgcgggtact aagaccccgg ttgcgaaaaa cggtcaactg
     4201 tctatcaagg gcacccaact ggtgaatcgt gatggcaagg ctgttcaact gaaaggtatt
     4261 tcctctcatg gtctgcaatg gtacggtgag tacgttaaca aggattctct gaagtggctg
     4321 cgtgacgact ggggcattac tgtgtttcgc gcggctatgt acaccgcgga cggtggctac
     4381 atcgataatc caagcgttaa gaacaaggtt aaggaggctg tggaggctgc gaaggaactg
     4441 ggtatctacg ttattatcga ttggcacatt ctgaacgatg gcaacccgaa tcagaacaag
     4501 gaaaaagcga aggagttttt caaagagatg agcagcctgt atggtaatac cccgaatgtt
     4561 atctacgaga tcgcgaatga gccgaatggc gacgttaatt ggaaacgcga catcaagccg
     4621 tacgctgagg aggttatcag cgtgattcgt aagaacgacc cggataacat tattatcgtt
     4681 ggtactggca cctggagcca agacgttaac gacgcggcgg acgatcaact gaaggacgcg
     4741 aatgtgatgt atgcgctgca cttctacgcg ggcacccacg gtcagttcct gcgtgacaag
     4801 gcgaattacg cgctgtctaa gggtgcgccg atcttcgtta ccgaatgggg cacctctgat
     4861 gcttccggta acggtggcgt ttttctggac cagagccgtg agtggctgaa gtacctggat
     4921 tctaaaacta tttcttgggt taattggaac ctgagcgaca aacaagagag cagcagcgcg
     4981 ctgaaaccag gtgcgagcaa gaccggtggt tggcgtctga gcgacctgag cgcgagcggt
     5041 acgtttgtgc gtgagaatat cctgggtact aaggacagca cctaaggatc caaactcgag
     5101 taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     5161 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     5221 ggcctttctg cgtttatacc tagggtacgg gttttgctgc ccgcaaacgg gctgttctgg
     5281 tgttgctagt ttgttatcag aatcgcagat ccggcttcag ccggtttgcc ggctgaaagc
     5341 gctatttctt ccagaattgc catgattttt tccccacggg aggcgtcact ggctcccgtg
     5401 ttgtcggcag ctttgattcg ataagcagca tcgcctgttt caggctgtct atgtgtgact
     5461 gttgagctgt aacaagttgt ctcaggtgtt caatttcatg ttctagttgc tttgttttac
     5521 tggtttcacc tgttctatta ggtgttacat gctgttcatc tgttacattg tcgatctgtt
     5581 catggtgaac agctttgaat gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt
     5641 ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatgtaa cggtgaacag
     5701 ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga gccataagaa
     5761 cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg tttttgcgtg
     5821 agccatgaga acgaaccatt gagatcatac ttactttgca tgtcactcaa aaattttgcc
     5881 tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg
     5941 ttatgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc
     6001 tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc
     6061 aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt
     6121 aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta
     6181 ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga
     6241 gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt
     6301 tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa
     6361 ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt
     6421 gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc
     6481 atcagctctc tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc
     6541 atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca
     6601 atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag
     6661 tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc
     6721 aattggtcta ggtgatttta atcactatac caattgagat gggctagtca atgataatta
     6781 ctagtccttt tcccgggtga tctgggtatc tgtaaattct gctagacctt tgctggaaaa
     6841 cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt
     6901 tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata
     6961 gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg
     7021 tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc
     7081 accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct
     7141 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg
     7201 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca
     7261 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat
     7321 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat
     7381 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
     7441 caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa aacgcccggc
     7501 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     7561 acaggagtcc aagcgagctc gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7621 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7681 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7741 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7801 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7861 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7921 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7981 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8041 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8101 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8161 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8221 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8281 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8341 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8401 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8461 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8521 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8581 gtgccacct
//
