LOCUS       sequence-60302-e        3026 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 3026)
  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 3026)
  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-UAS1B2-Leum
FEATURES             Location/Qualifiers
     source          1..3026
                     /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(805..821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(805..821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(818..840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    829..845
                     /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(853..859,860..877,878..883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    898..919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1036..1053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1207..1795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1287..1306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1966..2757,2758..2826))
                     /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     2589..2608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2827..2931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2999..3017
                     /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 tcgagctcgg tacccgggga tccctgaggt gtctcacaag tgccgtgcag tcccgccccc
      481 acttgcttct ctttgtgtgt agtgtacgta cattatcgag accgttgttc ccgcccacct
      541 cgatccgggt cgacctgagg tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc
      601 tctttgtgtg tagtgtacgt acattatcga gaccgttgtt cccgcccacc tcgatccggg
      661 catgcactga tcacgggcaa aagtgcgtat atatacaaga gcgtttgcca gccacagatt
      721 ttcactccac acaccacatc acacatacaa ccacacacat ccacaatgga acccgaaact
      781 aaaagcttca ggcgcgccat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1621 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     1681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2941 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3001 taggcgtatc acgaggccct ttcgtc
//
