LOCUS       sequence-254847-        2643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 destination vector for a promoter with A - B scars. Clone in
            with BpiI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2643)
  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 2643)
  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: pEMY07AB-PRO
FEATURES             Location/Qualifiers
     source          1..2643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      66..654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     555..574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    884..905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(920..925,926..943,944..950)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    958..974
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     963..985
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     982..998
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     982..998
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             994..1332
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSILEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRRPALRP
                     STICSDAA"
     misc_feature    1011..1067
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1068..1085)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1068..1084)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1077..1099)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1308..1327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1427..1449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1487..1505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1573..1677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1678..1746,1747..2538)
                     /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     complement(1896..1915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
       61 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg gctcaagacg atagttaccg
      361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      781 gctcggtctc tcattaggtc ttccccgcgc gttggccgat tcattaatgc agctggcacg
      841 acaggtttcc ctactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      901 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      961 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
     1021 gcctgcaggt cgattctaga ggatccccgg gtaccgagct cgaattcact ggccgtcgtt
     1081 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     1141 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1201 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     1261 ggtatttcac accgcatatg gtgcagaaga cctgcactga gacccagtac aatctgctct
     1321 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1381 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1441 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     1501 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc
     2401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2461 acggggagcc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2641 aaa
//
