LOCUS       sequence-253027-        3503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 destination vector for a transcription unit, pathway 
            position 7. Clone a transcription unit in with BsaI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3503)
  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 3503)
  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-f3
FEATURES             Location/Qualifiers
     source          1..3503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..20
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     118..135
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     119..135
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    605..638
                     /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             complement(710..1015)
                     /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     complement(812..831)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1415..1448)
                     /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(1655..1674)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1741..1832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1833..2642
                     /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"
     rep_origin      2764..3352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3253..3272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagcgagtc agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac
       61 taagcagaag gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt
      121 aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt
      181 aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga
      241 aagagcattt gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat
      301 ttaaccttat aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat
      361 tgatgaacac ctataattaa actattcatc tattatttat gattttttgt atatacaata
      421 tttctagttt gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca
      481 gtttttgata tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag
      541 atgttatcag tatttattat gcatttagaa taaattttgt gtcgccctta ttcgactcac
      601 tatagaagtt cctattctct agaaagtata ggaacttcac ttcattttac ttagtatggt
      661 ctgttggaaa ggattgtggc ttcgcataca ggctttctga ggcgagacct tatattcccc
      721 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
      781 cttccccgat aacggaaacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
      841 catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg
      901 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
      961 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
     1021 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
     1081 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
     1141 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
     1201 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
     1261 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatatgcata
     1321 ctgttatctg gcttttagta agccggtctc cgcacgtgca ctacatcttg tgggtgtcga
     1381 catgctggct ggattcgaag gtttccccac ttcagaagtt cctatacttt ctagagaata
     1441 ggaacttcac tatagagtcg aataagggcg acacccccta attagcccgg gcgaaaggcc
     1501 cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg
     1561 gggagtcccc acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc
     1621 aggtgggacc accgcgctac tgccgccagg caaacaaggg gtgttatgag ccatattcag
     1681 gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta
     1741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1801 aaatgcttca ataatattga aaaaggaaga atatgagcca tattcaacgg gaaacgtcga
     1861 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     1921 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     1981 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2041 taaactggct gacggaattt atgccacttc cgaccatcaa gcattttatc cgtactcctg
     2101 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcgttccag gtattagaag
     2161 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     2221 actcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc ctcgctcagg
     2281 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     2341 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     2401 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     2461 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     2521 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     2581 gtattgataa tcctgatatg aataaattgc aatttcattt gatgctcgat gagtttttct
     2641 aagcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2701 gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca
     2941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3481 tcgccgcagc cgaacgaccg agc
//
