LOCUS       sequence-60364-e        5035 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 5035)
  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 5035)
  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-UAS1B20-Leum
FEATURES             Location/Qualifiers
     source          1..5035
                     /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(2814..2830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2814..2830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2827..2849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2838..2854
                     /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(2862..2868,2869..2886,2887..2892))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2907..2928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3045..3062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3216..3804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3296..3315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3975..4766,4767..4835))
                     /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     4598..4617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4836..4940)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5008..5026
                     /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 tggtacccga
     2221 attcgagctc ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt
     2281 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccggtctag
     2341 actgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag
     2401 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccggggat ccctgaggtg
     2461 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac
     2521 attatcgaga ccgttgttcc cgcccacctc gatccgggtc gacctgaggt gtctcacaag
     2581 tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt agtgtacgta cattatcgag
     2641 accgttgttc ccgcccacct cgatccgggc atgcactgat cacgggcaaa agtgcgtata
     2701 tatacaagag cgtttgccag ccacagattt tcactccaca caccacatca cacatacaac
     2761 cacacacatc cacaatggaa cccgaaacta aaagcttcag gcgcgccatc atggtcatag
     2821 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2881 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2941 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3001 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3061 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4261 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4561 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4681 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4981 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
