LOCUS       sequence-60306-e        3449 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 3449)
  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 3449)
  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-UAS1B6-Leum
FEATURES             Location/Qualifiers
     source          1..3449
                     /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(1228..1244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1228..1244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1241..1263)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1252..1268
                     /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(1276..1282,1283..1300,1301..1306))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1321..1342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1459..1476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1630..2218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1710..1729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2389..3180,3181..3249))
                     /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     3012..3031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3250..3354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3422..3440
                     /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 ggtcgacctg aggtgtctca caagtgccgt gcagtcccgc ccccacttgc
     1021 ttctctttgt gtgtagtgta cgtacattat cgagaccgtt gttcccgccc acctcgatcc
     1081 gggcatgcac tgatcacggg caaaagtgcg tatatataca agagcgtttg ccagccacag
     1141 attttcactc cacacaccac atcacacata caaccacaca catccacaat ggaacccgaa
     1201 actaaaagct tcaggcgcgc catcatggtc atagctgttt cctgtgtgaa attgttatcc
     1261 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1321 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1381 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1441 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1501 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1561 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2041 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     2101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2341 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2401 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2461 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2521 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2581 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2641 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2701 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2761 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2821 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2881 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2941 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3001 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3061 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3121 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3181 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3241 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3361 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3421 aaataggcgt atcacgaggc cctttcgtc
//
