LOCUS       sequence-233259-        7088 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses IUP pathway genes (ChK, IPK, idi) under constitutive 
            promoter. Kan. resist..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7088)
  AUTHORS   Chatzivasileiou AO, Ward V, Edgar SM, Stephanopoulos G
  TITLE     Two-step pathway for isoprenoid synthesis.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Dec 24. pii: 1812935116. doi: 
            10.1073/pnas.1812935116.
  PUBMED    30584096
REFERENCE   2  (bases 1 to 7088)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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. 2018 Dec 24. pii: 1812935116. doi: 
            10.1073/pnas.1812935116."
COMMENT     Sequence Label: pSEVA228-pro4IUPi
FEATURES             Location/Qualifiers
     source          1..7088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1143..1151
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             2158..2166
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(2760..2778)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2774..2821
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2958..2977)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2969..3784
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            3951..4059
                     /note="incP origin of transfer"
     CDS             complement(4120..5268)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      5661..6192
                     /label=oriV
                     /note="incP origin of replication"
     RBS             6448..6456
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 tcgtgaatgg gcgcttcgag cagttcctcg aaaactccaa aacgctgacc aaggatgata
       61 tccggaactg gaaaaactcg caacggatcg cccgccgcat gaaggagctg catgtgggcg
      121 tgcccctcct ctcgtcggag cggaagaatg ggagcgcctg ctggcaaaaa atcaaccaat
      181 ggctccgcac gatcgagaag gtggatcagt gggtcgggga cccgaagaac atcgagaaca
      241 gcctcctctg cgaaaattgg tccaaattca tggacatcgt cgatcggtac cacaagtggc
      301 tgatcagcca agaacaaggg atcgagcaag tcaacaaaaa tctgatcttc tgccataatg
      361 atgcccaata cgggaatctc ctcttcaccg cgcccgtcat gaacaccccc tccctgtata
      421 ccgcgccgag ctcgacctcc ctgacgtccc aaagcagcag cctcttcccc tcgtccagca
      481 acgtgatcgt cgatgatatc atcaatcccc cgaagcaaga acaatcccaa gattccaaac
      541 tcgtggtcat cgatttcgaa tacgccgggg ccaatcccgc cgcgtacgat ctcgccaatc
      601 acctctcgga atggatgtac gactataata acgccaaagc cccgcaccag tgccacgccg
      661 accggtaccc cgacaaggag caagtgctca acttcctgta ttcgtatgtc agccatctcc
      721 gcggcggggc caaagagccc atcgatgaag aagtccagcg cctctataaa tcgatcatcc
      781 agtggcgccc cacggtgcag ctcttctggt cgctgtgggc gatcctgcaa agcggcaagc
      841 tggaaaaaaa agaagccagc accgccatca cccgcgaaga aatcgggccc aatgggaaaa
      901 agtatatcat caagacggag cccgagtcgc ccgaagagga cttcgtcgaa aatgacgacg
      961 aacccgaagc cggcgtgtcg atcgatacct tcgactacat ggcctacggg cgggacaaga
     1021 tcgcggtgtt ctggggggac ctgatcgggc tgggcatcat cacggaggag gaatgcaaga
     1081 acttctcgag cttcaaattc ctcgacacca gctacctgta agtacaagaa aagtcagtag
     1141 tctaaggagg taagcatgga actcaatatc agcgaaagcc ggtcgcgcag catccggtgc
     1201 atcgtgaagc tcgggggcgc ggccatcacc tgcaaaaacg aactcgaaaa gatccatgac
     1261 gaaaacctcg aagtggtggc ctgccaactg cggcaagcga tgctggaagg ctccgccccc
     1321 tccaaagtca tcggcatgga ctggtccaaa cggccgggct cctccgaaat ctcctgcgat
     1381 gtggacgaca tcggcgacca gaaatcgagc gaattctcga agttcgtggt cgtccacggg
     1441 gcgggctcgt tcggccattt ccaagcgtcg cggtcgggcg tccataaagg gggcctggag
     1501 aagcccatcg tcaaagccgg gttcgtcgcg acccggatct cggtcaccaa tctcaatctc
     1561 gagatcgtcc gcgccctggc ccgggaaggc atcccgacga tcgggatgag ccccttcagc
     1621 tgcggctggt ccacctccaa acgggatgtc gcgtcggcgg atctcgcgac ggtcgccaag
     1681 accatcgact cgggcttcgt gcccgtgctc catggggacg cggtcctgga caacatcctg
     1741 ggctgcacga tcctgtccgg cgatgtcatc atccggcacc tggccgacca tctcaagccg
     1801 gaatacgtcg tgttcctgac cgacgtgctc ggggtctatg accggccccc gagcccgtcg
     1861 gagccggacg ccgtcctgct caaggagatc gccgtcgggg aggatggctc ctggaaggtg
     1921 gtcaaccccc tcctggaaca taccgacaaa aaggtggatt attccgtcgc ggcccacgat
     1981 accacggggg ggatggaaac caaaatctcg gaagccgcca tgatcgccaa gctcggcgtg
     2041 gatgtgtaca tcgtgaaagc cgcgaccacc cattcgcagc gggcgctcaa tggcgacctg
     2101 cgcgactccg tccccgaaga ctggctgggg accatcatcc gcttcagcaa ataaaactaa
     2161 ggaggtctat atgcaaacgg aacacgtcat tttattgaat gcacagggag ttcccacggg
     2221 tacgctggaa aagtatgccg cacacacggc agacacccgc ttacatctcg cgttctccag
     2281 ttggctgttt aatgccaaag gacaattatt agttacccgc cgcgcactga gcaaaaaagc
     2341 atggcctggc gtgtggacta actcggtttg tgggcaccca caactgggag aaagcaacga
     2401 agacgcagtg atccgccgtt gccgttatga gcttggcgtg gaaattacgc ctcctgaatc
     2461 tatctatcct gactttcgct accgcgccac cgatccgagt ggcattgtgg aaaatgaagt
     2521 gtgtccggta tttgccgcac gcaccactag tgcgttacag atcaatgatg atgaagtgat
     2581 ggattatcaa tggtgtgatt tagcagatgt attacacggt attgatgcca cgccgtgggc
     2641 gttcagtccg tggatggtga tgcaggcgac aaatcgcgaa gccagaaaac gattatctgc
     2701 atttacccag cttaaataag ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2761 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2821 gctgaaagga ggaactatat cgggtcccca ataattacga tttaaatttg tgtctcaaaa
     2881 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2941 ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa cgagctgtag
     3001 ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat gggcgcgtga
     3061 taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg atgcgccgga
     3121 actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg aaatggtgcg
     3181 tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta ttcgcacccc
     3241 ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc aggtgctgga
     3301 agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc tgcgtcgtct
     3361 gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc gtctggccca
     3421 ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg atgaacgtaa
     3481 cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt ttagcccgga
     3541 tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg aaggcaaact
     3601 gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg atctggccat
     3661 tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt ttcagaaata
     3721 tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg atgaattttt
     3781 ctaataatta attggaccgc ggtccgcgcg ttgtcctttt ccgctgcata accctgcttc
     3841 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
     3901 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
     3961 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
     4021 gcggtgctca acgggaatcc tgctctgcga ggctggccgt aggccggccg cgatgcaggt
     4081 ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag
     4141 gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc
     4201 gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa
     4261 ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg
     4321 ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc
     4381 aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg
     4441 gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga
     4501 cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata
     4561 ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg
     4621 ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc
     4681 gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat
     4741 gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga
     4801 gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa
     4861 ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc
     4921 ttcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt
     4981 tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg
     5041 cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc
     5101 gcgctcgatc ttggccgtag cttgctggac tatcgagccg acggactgga aggtttcgcg
     5161 gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc
     5221 gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg
     5281 cgggattgcc ccggaattaa ttccccggat cgatccgtcg atcttgatcc cctgcgccat
     5341 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca
     5401 gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc
     5461 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt
     5521 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     5581 ttctacgtgg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg
     5641 ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt
     5701 cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat
     5761 tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt
     5821 gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca
     5881 tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc
     5941 gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg
     6001 gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc
     6061 cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca
     6121 agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca
     6181 acgccggcgg ccctacatgg ctctgctgta gtgagtgggt tgcgctccgg cagcggtcct
     6241 gatcccccgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggcgcgccc
     6301 agctgttcta gagcacagct aacaccacgt cgtccctatc tgctgcccta ggtctatgag
     6361 tggttgctgg ataactttac gggcatgcat aaggctcgga tgatatattc agggagacca
     6421 caacggtttc aacgcaagga aacacattaa ggaggtttaa atggtgcagg agtcccgccc
     6481 cggctcggtc cggtcgtatt ccgtgggcta ccaggcccgg tcgcggtcgt cgtcccagcg
     6541 ccgccattcg ctcacgcggc agcgcagcag ccagcggctc atccggacga tctccatcga
     6601 gagcgatgtg agcaatatca cggacgatga tgatctgcgg gcggtgaatg aaggggtggc
     6661 cggggtccag ctcgacgtct ccgagacggc gaacaaaggg ccacgccggg ccagtgccac
     6721 cgatgtcacc gactcgctgg gctccacgtc cagcgaatat atcgagatcc ccttcgtgaa
     6781 agagacgctg gacgcgagcc tcccctcgga ttacctcaaa caagacatcc tgaacctgat
     6841 ccaatccctg aagatctcga aatggtacaa taacaaaaag atccagcccg tcgcccagga
     6901 catgaacctc gtcaaaatct ccggcgcgat gaccaatgcg atcttcaagg tggagtaccc
     6961 gaaactgccg tccctcctgc tgcggatcta tggcccgaat atcgataaca tcatcgaccg
     7021 cgaatatgaa ctccagatcc tcgcgcggct ctcgctgaaa aacatcgggc cgtccctgta
     7081 cggctgct
//
