LOCUS       sequence-46430-e        7102 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 7102)
  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 7102)
  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: pXylan
FEATURES             Location/Qualifiers
     source          1..7102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1509..1540)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      3631..3702
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3718..3745
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4276..5226)
                     /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(5988..6082)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(6112..6903,6904..6972))
                     /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     6735..6754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6973..7077)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg aacccgacag aattagatga
       61 gattgcagga aaactcggtt aacggagtga tcgagttaac attgttaagt taaatattgg
      121 tttcaactcc gatttacatg gttgctgtgt tgttaaattg tacaaagatg ttatagaaac
      181 aaaatgtaac atctctatgg acacgcacac ggataacaac tgaattcaaa agatctagga
      241 ggaaaaaaaa atgtctacag aaaatgaagt tgttgattac aaagcgctgg cggccagggc
      301 gatttcccaa cccctggtaa cccatatcta tacggctgac ccctcagcac atgtattcaa
      361 tggcaaggtt tacatttatc catcacacga tatcgatgcc ggtatcccgt tcaatgacaa
      421 cggtgaccac ttcgggatgg aggactacca tgtgttgcgt atggattccc ctgaaggcaa
      481 ggcagaagac tgtggtgtgg cactgcatgt gaaggatgtg ccctgggcag agcgtcaaat
      541 gtgggcgccg gacgcaatca ccaaagatgg caagtactac ctctatttcc ccgctcgcgc
      601 ccgtgatgga ctgtttaaaa tcggggttgc gataggtgat caacctgaag ggccttttgt
      661 tgcagagcct gagcctatcg caggaagcta ttccattgat cctgcggtct ttggcgatga
      721 tgacggtgag ttttacctgt acttcggcgg tatttggggc gggcaattac aaaaataccg
      781 cgataacact tacagcgaaa tccatgagga gcccacggcc gaccaacccg cactgggcgc
      841 ccgtgtagcg cgcctgagtg ctgatatgaa atcctttgtg gaagcttcgc gcgaagtggt
      901 aattctggat gagcagggcc agccgctatt ggcgggcgat aacagtcgcc gttattttga
      961 aggtccctgg atgcacaaat accagggcaa gtactacctg tcctactcaa ccggtgatac
     1021 tcacttcttg tgttatgcca ccagtgataa tccctatggg cctttcacct atcaagggca
     1081 gattttgaca ccggttgtgg gctggacaac acaccattcg atctgtgaat tcgaaggcaa
     1141 gtggtatttg ttttaccacg actctgtctt gtcggaaggg gtgacacatt tgcgttcggt
     1201 gaaggtgaca gaattgcact atgaggcgga tgggaaaatt aaaacgattc atccctatcg
     1261 tgattagtaa gtatggtcga cgatccggca aacaaaccac cgttggtagc ggtggttttt
     1321 ttgtttggat cgacaatctt cgtaagcgtc atcaataagc gtaaaaaaac cgggcaatgc
     1381 ccggtttttt aatgagaaat tttacctgtc gtagccgcca ccatccggca aagaagcata
     1441 caaggctttt ggcttatagc tacgtagcgc attgcgtcgc agcacaatcc cggcaccgat
     1501 cgagtaatag tcaaaagcct ccggtcggag gcttttgact ttctgccctt gtcagttatc
     1561 atcttcggtt acggttatcg gcagaacgcc ctgccgctga caagagctta tttgcctgac
     1621 caatctactc cacatgatag ataactatca tctatcgttg gcatcagcga catctgtcac
     1681 attcctgtca atagcgttaa ctgcttcaaa tttttgattc atacctgccc cttgtcccct
     1741 gtcaaatgct tgacggctcg ccctaattct ctaaattgta tttctagagt tggcgaggtt
     1801 ttgaacagcc ccctctctga ccccggttta ttccatctta cttgtataag atttgcgaag
     1861 gatgtcgaag cgaattcaaa agatctagca ggaggaaaaa aaaatgacta tgacaagact
     1921 gaagatttcg aaaactctgc tggctgtaat gttgacctct gccgtcgcga ccggctctgc
     1981 ctacgcggaa aacaacgcgc agactaccaa tgaaagcgca gggcaaaaag tcgatagctc
     2041 tatgaataaa gtcggtaatt tcatggatga cagcgccatc accgcgaaag tgaaggcggc
     2101 cctggtggat catgacaaca tcaagagcac cgatatctct gtaaaaaccg atcaaaaagt
     2161 cgtgaccctg agcggtttcg ttgaaagcca ggcccaggcc gaagaggcag tgaaagtggc
     2221 gaaaggcgtt gaaggggtga cctctgtcag cgacaaactg cacgttcgcg acgctaaaga
     2281 aggctcggtg aagggctacg cgggtgacac cgccaccacc agtgaaatca aagccaaact
     2341 gctggcggac gatatcgtcc cttcccgtca tgtgaaagtt gaaaccaccg acggcgtggt
     2401 tcagctctcc ggtaccgtcg attctcaggc acaaagtgac cgtgctgaaa gtatcgccaa
     2461 agcggtagat ggtgtgaaaa gcgttaaaaa tgatctgaaa actaagggca gcggatctat
     2521 gaccggtaag aaagcattca acgatcagac ctccgcagaa gatatcccaa gcctggcgga
     2581 ggcttttcgc gactattttc cgatcggtgc ggcgatcgaa ccgggttata ccactggtca
     2641 gattgctgaa ctgtataaga aacatgttaa tatgctggtg gccgagaacg ctatgaaacc
     2701 ggcgtctctg cagccgaccg aaggtaactt ccagtgggcg gatgcagatc gtatcgttca
     2761 gtttgctaaa gaaaacggca tggaactgcg cttccacacc ctggtttggc ataaccaaac
     2821 cccgactggt ttctccctgg acaaagaagg taaaccgatg gtggaagaaa ccgacccgca
     2881 gaaacgtgaa gagaaccgta aactgctgct gcagcgtctg gagaactaca tccgtgcagt
     2941 tgtgctgcgt tataaagatg acatcaaatc ttgggatgta gtgaacgaag ttattgaacc
     3001 aaacgatcca ggtggtatgc gtaactctcc ttggtaccag atcactggta ctgaatacat
     3061 cgaggtcgct ttccgcgcag cacgtgaagc cggtggttcc gacatcaaac tgtacatcaa
     3121 cgactataac accgatgatc cggttaaacg tgacatcctg tatgagctgg ttaaaaacct
     3181 gctggaaaaa ggcgtaccta tcgacggtgt tggtcaccag acccatattg atatctataa
     3241 tccgccggta gaacgcatca ttgaatccat taaaaagttc gcgggcctgg gcctggacaa
     3301 catcatcacc gaactggaca tgagcattta ctcctggaac gaccgttccg attatggtga
     3361 ctctattccg gattacattc tgaccctgca ggcaaagcgt tatcaggagc tgttcgatgc
     3421 gctgaaagaa aacaaagaca ttgtttccgc ggtcgtgttc tggggcattt ctgacaaata
     3481 ctcttggctg aacggcttcc cggtgaagcg caccaacgcc cctctgctgt ttgatcgtaa
     3541 ctttatgccg aaacctgcct tctgggcaat cgtggaccca tcccgtctgc gtgaataagg
     3601 atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac
     3661 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     3721 tcaccttcgg gtgggccttt ctgcgtttat acctagggta cgggttttgc tgcccgcaaa
     3781 cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt cagccggttt
     3841 gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac gggaggcgtc
     3901 actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg tttcaggctg
     3961 tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc atgttctagt
     4021 tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc atctgttaca
     4081 ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa aagctctgat
     4141 gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc ctttgatatg
     4201 taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca
     4261 agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
     4321 ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact
     4381 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
     4441 ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
     4501 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
     4561 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
     4621 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
     4681 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
     4741 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     4801 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     4861 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     4921 cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
     4981 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     5041 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
     5101 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     5161 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     5221 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     5281 tcaatgataa ttactagtcc ttttcccggg tgatctgggt atctgtaaat tctgctagac
     5341 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     5401 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     5461 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     5521 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     5581 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
     5641 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
     5701 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
     5761 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
     5821 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
     5881 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     5941 gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata
     6001 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     6061 actggatcta tcaacaggag tccaagcgag ctcgtaaact tggtctgaca gttaccaatg
     6121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7081 atttccccga aaagtgccac ct
//
