LOCUS       sequence-253021-        3504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 destination vector for a transcription unit, pathway 
            position 2. Clone a transcription unit in with BsaI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3504)
  AUTHORS   Young EM, Zhao Z, Gielesen BEM, Wu L, Benjamin Gordon D, Roubos JA, 
            Voigt CA
  TITLE     Iterative algorithm-guided design of massive strain libraries, 
            applied to itaconic acid production in yeast.
  JOURNAL   Metab Eng. 2018 Jul;48:33-43. doi: 10.1016/j.ymben.2018.05.002. Epub
            2018 May 9.
  PUBMED    29753070
REFERENCE   2  (bases 1 to 3504)
  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: "Metab 
            Eng."; date: "2018-07"; volume: "48:33-43. doi"; pages: " 
            10.1016/j.ymben.2018.05.002. Epub 2018 May 9"
COMMENT     Sequence Label: pDSM-ab
FEATURES             Location/Qualifiers
     source          1..3504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    233..266
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             667..972
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     851..870
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1044..1077)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1547..1564)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1547..1563)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1662..1679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1833..2421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1913..1932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2543..3352)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3353..3444)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg ctcataacac cccttgtttg cctggcggca gtagcgcggt ggtcccacct
       61 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggactccc
      121 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
      181 ggcctttcgc ccgggctaat tagggggtgt cgcccttatt cgactctata gtgaagttcc
      241 tattctctag aaagtatagg aacttctgaa gtggggttgc ccatcgaacg tacaagtact
      301 cctctgttct ctccttcctt tgctttgtgc ggagaccggc ttactaaaag ccagataaca
      361 gtatgcatat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
      421 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
      481 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
      541 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
      601 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
      661 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
      721 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
      781 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
      841 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtttccgt tatcggggaa
      901 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
      961 tggggaatat aaggtctcgc ctccggatcg atgtacacaa ccgactgcac ccaaacgaac
     1021 acaaatctta gcaaaaatga agtgaagttc ctatactttc tagagaatag gaacttctat
     1081 agtgagtcga ataagggcga cacaaaattt attctaaatg cataataaat actgataaca
     1141 tcttatagtt tgtattatat tttgtattat cgttgacatg tataattttg atatcaaaaa
     1201 ctgattttcc ctttattatt ttcgagattt attttcttaa ttctctttaa caaactagaa
     1261 atattgtata tacaaaaaat cataaataat agatgaatag tttaattata ggtgttcatc
     1321 aatcgaaaaa gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt tataaggtta
     1381 aataattctc atatatcaag caaagtgaca ggcgccctta aatattctga caaatgctct
     1441 ttccctaaac tccccccata aaaaaacccg ccgaagcggg tttttacgtt atttgcggat
     1501 taacgattac tcgttatcag aaccgcccag ggggcccgag cttaagactg gccgtcgttt
     1561 tacaacacag aaagagtttg tagaaacgca aaaaggccat ccgtcagggg ccttctgctt
     1621 agtttgatgc ctggcagttt cctactctcg ccttccgctt cctcgctcac tgactcgctg
     1681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2221 ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag aagaacagta
     2281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2461 tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagcttg cgccgtcccg
     2521 tcaagtcagc gtaatgctct gcttagaaaa actcatcgag catcaaatga aattgcaatt
     2581 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     2641 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     2701 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     2761 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     2821 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     2881 aaccgttatt cattcgtgat tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag
     2941 gacaattaca aacaggaatc gagtgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     3001 tattttcacc tgaatcagga tattcttcta atacctggaa cgctgttttt ccggggatcg
     3061 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagtg
     3121 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     3181 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga
     3241 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     3301 ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc atattcttcc
     3361 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3421 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     3481 acattatcgc gagcccattt atac
//
