LOCUS       sequence-60311-e        4026 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 4026)
  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 4026)
  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-UAS1B11-Leum
FEATURES             Location/Qualifiers
     source          1..4026
                     /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(1805..1821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1805..1821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1818..1840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1829..1845
                     /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(1853..1859,1860..1877,1878..1883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1898..1919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2036..2053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2207..2795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2287..2306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2966..3757,3758..3826))
                     /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     3589..3608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3827..3931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3999..4017
                     /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 aattcctgag
      421 gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg
      481 tacattatcg agaccgttgt tcccgcccac ctcgatccgg tctagactga ggtgtctcac
      541 aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc
      601 gagaccgttg ttcccgccca cctcgatccg gggatccctg aggtgtctca caagtgccgt
      661 gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt
      721 gttcccgccc acctcgatcc gggtcgacct gaggtgtctc acaagtgccg tgcagtcccg
      781 cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc
      841 cacctcgatc cgggagctcc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt
      901 gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat
      961 ccggtctaga ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt
     1021 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccggggatc
     1081 cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag
     1141 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccgggtcg acctgaggtg
     1201 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac
     1261 attatcgaga ccgttgttcc cgcccacctc gatccgggca tgcggtaccc gaattcgagc
     1321 tcctactcta gactgaggtg tctcacaagt gccgtgcagt cccgccccca cttgcttctc
     1381 tttgtgtgta gtgtacgtac attatcgaga ccgttgttcc cgcccacctc gatccgggga
     1441 tccctgaggt gtctcacaag tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt
     1501 agtgtacgta cattatcgag accgttgttc ccgcccacct cgatccgggt cgacctgagg
     1561 tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt
     1621 acattatcga gaccgttgtt cccgcccacc tcgatccggg catgcactga tcacgggcaa
     1681 aagtgcgtat atatacaaga gcgtttgcca gccacagatt ttcactccac acaccacatc
     1741 acacatacaa ccacacacat ccacaatgga acccgaaact aaaagcttca ggcgcgccat
     1801 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1861 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1921 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1981 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2041 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2101 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2161 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2221 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2281 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2341 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2401 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2461 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2521 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2581 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2641 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2701 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2761 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2821 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2881 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2941 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3001 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3061 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3121 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3181 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3241 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3301 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3361 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3421 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3481 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3541 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3601 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3661 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3721 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3781 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3841 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3901 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3961 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4021 ttcgtc
//
