LOCUS       sequence-60313-e        4232 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 4232)
  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 4232)
  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-UAS1B13-Leum
FEATURES             Location/Qualifiers
     source          1..4232
                     /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(2011..2027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2011..2027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2024..2046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2035..2051
                     /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(2059..2065,2066..2083,2084..2089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2104..2125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2242..2259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2413..3001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2493..2512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3172..3963,3964..4032))
                     /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     3795..3814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4033..4137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4205..4223
                     /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 gaattcctga
     1321 ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac
     1381 gtacattatc gagaccgttg ttcccgccca cctcgatccg gtctagactg aggtgtctca
     1441 caagtgccgt gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat
     1501 cgagaccgtt gttcccgccc acctcgatcc ggggatccct gaggtgtctc acaagtgccg
     1561 tgcagtcccg cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt
     1621 tgttcccgcc cacctcgatc cgggtcgacc tgaggtgtct cacaagtgcc gtgcagtccc
     1681 gcccccactt gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc
     1741 ccacctcgat ccgggagctc ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact
     1801 tgcttctctt tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga
     1861 tccgggcatg cactgatcac gggcaaaagt gcgtatatat acaagagcgt ttgccagcca
     1921 cagattttca ctccacacac cacatcacac atacaaccac acacatccac aatggaaccc
     1981 gaaactaaaa gcttcaggcg cgccatcatg gtcatagctg tttcctgtgt gaaattgtta
     2041 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2161 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2821 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4201 taaaaatagg cgtatcacga ggccctttcg tc
//
