LOCUS       sequence-60305-e        3338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3338)
  AUTHORS   Blazeck J, Liu L, Redden H, Alper H
  TITLE     Tuning gene expression in Yarrowia lipolytica by a hybrid promoter 
            approach.
  JOURNAL   Appl Environ Microbiol. 2011 Nov;77(22):7905-14. doi: 
            10.1128/AEM.05763-11. Epub 2011 Sep 16.
  PUBMED    21926196
REFERENCE   2  (bases 1 to 3338)
  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"; doi: 
            "10.1128/AEM.05763-11"; journalName: "Appl Environ Microbiol"; date:
            "2011-11"; volume: "77"; issue: "22"; pages: "7905-14"
COMMENT     Sequence Label: pUC-UAS1B5-Leum
FEATURES             Location/Qualifiers
     source          1..3338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1117..1133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1117..1133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1130..1152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1141..1157
                     /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)."
     promoter        complement(join(1165..1171,1172..1189,1190..1195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1210..1231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1348..1365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1519..2107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1599..1618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2278..3069,3070..3138))
                     /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     2901..2920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3139..3243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3311..3329
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtttcga actgcagttg gtacccgaat
      421 tcctgaggtg tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta
      481 gtgtacgtac attatcgaga ccgttgttcc cgcccacctc gatccggtct agactgaggt
      541 gtctcacaag tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt agtgtacgta
      601 cattatcgag accgttgttc ccgcccacct cgatccgggg atccctgagg tgtctcacaa
      661 gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt acattatcga
      721 gaccgttgtt cccgcccacc tcgatccggg tcgacctgag gtgtctcaca agtgccgtgc
      781 agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg tacattatcg agaccgttgt
      841 tcccgcccac ctcgatccgg gagctcctga ggtgtctcac aagtgccgtg cagtcccgcc
      901 cccacttgct tctctttgtg tgtagtgtac gtacattatc gagaccgttg ttcccgccca
      961 cctcgatccg ggcatgcact gatcacgggc aaaagtgcgt atatatacaa gagcgtttgc
     1021 cagccacaga ttttcactcc acacaccaca tcacacatac aaccacacac atccacaatg
     1081 gaacccgaaa ctaaaagctt caggcgcgcc atcatggtca tagctgtttc ctgtgtgaaa
     1141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1921 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3241 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3301 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
