LOCUS       sequence-60366-e        5929 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 5929)
  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 5929)
  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-UAS1B28-Leum
FEATURES             Location/Qualifiers
     source          1..5929
                     /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(3708..3724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3708..3724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3721..3743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3732..3748
                     /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(3756..3762,3763..3780,3781..3786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3801..3822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3939..3956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4110..4698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4190..4209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4869..5660,5661..5729))
                     /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     5492..5511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5730..5834)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5902..5920
                     /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 aggtacccga attcctgagg
      421 tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt
      481 acattatcga gaccgttgtt cccgcccacc tcgatccggt ctagactgag gtgtctcaca
      541 agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg tacattatcg
      601 agaccgttgt tcccgcccac ctcgatccgg ggatccctga ggtgtctcac aagtgccgtg
      661 cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc gagaccgttg
      721 ttcccgccca cctcgatccg ggtcgacctg aggtgtctca caagtgccgt gcagtcccgc
      781 ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt gttcccgccc
      841 acctcgatcc gggagctcct gaggtgtctc acaagtgccg tgcagtcccg cccccacttg
      901 cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc cacctcgatc
      961 cggtctagac tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt gcttctcttt
     1021 gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat ccggggatcc
     1081 ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt
     1141 gtacgtacat tatcgagacc gttgttcccg cccacctcga tccgggtcga cctgaggtgt
     1201 ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag tgtacgtaca
     1261 ttatcgagac cgttgttccc gcccacctcg atccgggcat gcggtacccg aattcctgag
     1321 gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg
     1381 tacattatcg agaccgttgt tcccgcccac ctcgatccgg tctagactga ggtgtctcac
     1441 aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc
     1501 gagaccgttg ttcccgccca cctcgatccg gggatccctg aggtgtctca caagtgccgt
     1561 gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt
     1621 gttcccgccc acctcgatcc gggtcgacct gaggtgtctc acaagtgccg tgcagtcccg
     1681 cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc
     1741 cacctcgatc cgggagctcc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt
     1801 gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat
     1861 ccggtctaga ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt
     1921 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccggggatc
     1981 cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag
     2041 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccgggtcg acctgaggtg
     2101 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac
     2161 attatcgaga ccgttgttcc cgcccacctc gatccgggca tgcctgcagt tgaattcctg
     2221 aggtgtctca caagtgccgt gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta
     2281 cgtacattat cgagaccgtt gttcccgccc acctcgatcc ggtctagact gaggtgtctc
     2341 acaagtgccg tgcagtcccg cccccacttg cttctctttg tgtgtagtgt acgtacatta
     2401 tcgagaccgt tgttcccgcc cacctcgatc cggggatccc tgaggtgtct cacaagtgcc
     2461 gtgcagtccc gcccccactt gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg
     2521 ttgttcccgc ccacctcgat ccgggtcgac ctgaggtgtc tcacaagtgc cgtgcagtcc
     2581 cgcccccact tgcttctctt tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg
     2641 cccacctcga tccgggagct cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac
     2701 ttgcttctct ttgtgtgtag tgtacgtaca ttatcgagac cgttgttccc gcccacctcg
     2761 atccggtcta gactgaggtg tctcacaagt gccgtgcagt cccgccccca cttgcttctc
     2821 tttgtgtgta gtgtacgtac attatcgaga ccgttgttcc cgcccacctc gatccgggga
     2881 tccctgaggt gtctcacaag tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt
     2941 agtgtacgta cattatcgag accgttgttc ccgcccacct cgatccgggt cgacctgagg
     3001 tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt
     3061 acattatcga gaccgttgtt cccgcccacc tcgatccggg catgcggtac ccgaattcga
     3121 gctcctgagg tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg
     3181 tagtgtacgt acattatcga gaccgttgtt cccgcccacc tcgatccggt ctagactgag
     3241 gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg
     3301 tacattatcg agaccgttgt tcccgcccac ctcgatccgg ggatccctga ggtgtctcac
     3361 aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc
     3421 gagaccgttg ttcccgccca cctcgatccg ggtcgacctg aggtgtctca caagtgccgt
     3481 gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt
     3541 gttcccgccc acctcgatcc gggcatgcac tgatcacggg caaaagtgcg tatatataca
     3601 agagcgtttg ccagccacag attttcactc cacacaccac atcacacata caaccacaca
     3661 catccacaat ggaacccgaa actaaaagct tcaggcgcgc catcatggtc atagctgttt
     3721 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3781 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3841 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3901 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3961 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4021 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4081 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4141 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4201 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4261 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4321 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4381 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4441 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4501 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4561 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4621 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4681 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4741 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4801 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4861 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4921 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4981 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5041 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5101 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5161 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5221 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5281 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5341 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5401 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5461 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5521 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5581 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5641 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5701 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5761 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5821 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5881 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
