LOCUS       sequence-253010-        2643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 destination vector for a terminator with C - D 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: pEMY07CD-TER
FEATURES             Location/Qualifiers
     source          1..2643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    312..333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(348..353,354..371,372..378)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    386..402
                     /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     391..413
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     410..426
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     410..426
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             422..760
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSILEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRRPFMRP
                     STICSDAA"
     misc_feature    439..495
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(496..513)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(496..512)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(505..527)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(736..755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     855..877
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(915..933)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1001..1105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1106..1174,1175..1966)
                     /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(1324..1343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2137..2643,1..82)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(2626..2643,1..2)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
       61 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      181 tattaccgcc tttgagtgag ctgataccgc tcggtctctg aggaggtctt ccccgcgcgt
      241 tggccgattc attaatgcag ctggcacgac aggtttccct actggaaagc gggcagtgag
      301 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
      361 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
      421 tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg attctagagg atccccgggt
      481 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      541 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      601 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
      661 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcagaagacc
      721 ttttatgaga cccagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
      781 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      841 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      901 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
      961 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1021 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1081 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1141 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1201 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1261 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1321 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1381 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1441 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1501 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1561 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1621 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1681 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1741 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1801 atctggagcc ggtgagcgtg gctctcgcgg tatcattgca gcactggggc cagatggtaa
     1861 gccctcccgt atcgtagtta tctacacgac ggggagccag gcaactatgg atgaacgaaa
     1921 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1981 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2041 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2281 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2401 taccgggttg ggctcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2641 tct
//
