LOCUS       sequence-482090-        3646 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3646)
  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 3646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 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-UAS1B8-Leum
FEATURES             Location/Qualifiers
     source          1..3646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..71)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     171..190
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     384..406
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     398..415
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     399..415
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1445..1461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1445..1461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1458..1480)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1469..1485
                     /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(1493..1499,1500..1517,1518..1523))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1538..1559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1676..1693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1847..2435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1927..1946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2606..3397,3398..3466))
                     /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     3229..3248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3467..3571)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3639..3646,1..11)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
       61 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      121 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      181 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      241 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      301 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      361 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
      421 cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag
      481 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccggtcta gactgaggtg
      541 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac
      601 attatcgaga ccgttgttcc cgcccacctc gatccgggga tccctgaggt gtctcacaag
      661 tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt agtgtacgta cattatcgag
      721 accgttgttc ccgcccacct cgatccgggt cgacctgagg tgtctcacaa gtgccgtgca
      781 gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt acattatcga gaccgttgtt
      841 cccgcccacc tcgatccggg agctcctgag gtgtctcaca agtgccgtgc agtcccgccc
      901 ccacttgctt ctctttgtgt gtagtgtacg tacattatcg agaccgttgt tcccgcccac
      961 ctcgatccgg tctagactga ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct
     1021 tctctttgtg tgtagtgtac gtacattatc gagaccgttg ttcccgccca cctcgatccg
     1081 gggatccctg aggtgtctca caagtgccgt gcagtcccgc ccccacttgc ttctctttgt
     1141 gtgtagtgta cgtacattat cgagaccgtt gttcccgccc acctcgatcc gggtcgacct
     1201 gaggtgtctc acaagtgccg tgcagtcccg cccccacttg cttctctttg tgtgtagtgt
     1261 acgtacatta tcgagaccgt tgttcccgcc cacctcgatc cgggcatgca ctgatcacgg
     1321 gcaaaagtgc gtatatatac aagagcgttt gccagccaca gattttcact ccacacacca
     1381 catcacacat acaaccacac acatccacaa tggaacccga aactaaaagc ttggcgtaat
     1441 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1501 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1561 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1621 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1681 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1741 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1801 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1861 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1921 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1981 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2041 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2101 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2161 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2221 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2281 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2341 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2401 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2521 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2581 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2641 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2701 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2761 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2821 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2881 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2941 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3001 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3061 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3121 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3181 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3241 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3301 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3361 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3421 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3481 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3541 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3601 tctaagaaac cattattatc atgacattaa cctataaaaa taggcg
//
