LOCUS       sequence-60449-e        4356 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 4356)
  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 4356)
  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-UAS1B8-TEF(804)
FEATURES             Location/Qualifiers
     source          1..4356
                     /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(2135..2151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2135..2151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2148..2170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2159..2175
                     /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(2183..2189,2190..2207,2208..2213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2228..2249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2366..2383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2617..2636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3296..4087,4088..4156))
                     /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     3919..3938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4157..4261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4329..4347
                     /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 gcctgcagaa gcttttgcgt
     1321 ttcgctccca cactacaccg taataccacg tcactctcat tgcaggttac cctgcccgta
     1381 gtcgctcgat ccacctcctc cttctctcgt gtgtgcagca aagaggcaga gatggagccc
     1441 gtatggtgag ccgtagagtg gagcgagggg cgatcttaca acggctgtcg gcggatataa
     1501 acggtatttt cacaattgca ccccagccag accgatagcc ggccgcaatc cgccacccac
     1561 aaccgtctac ctcccacaga accccgtcac ttccaccctt ttccaccaga tcatatgtcc
     1621 caacttgcca aattaaaacc gtgcgaattt tcaaaataaa ctttggcaaa gaggctgcaa
     1681 aggaggggct ggtgagggcg tctggaagtc gaccagagac cgggttggcg gcgtatttgt
     1741 gtcccaaaaa acagccccaa ttgccccaat tgaccccaaa ttgacccagt agcgggccca
     1801 accccggcga gagccccctt caccccacat atcaaacctc ccccggttcc cacacttgcc
     1861 gttaagggcg tagggtactg cagtctggaa tctacgcttg ttcagacttt gtactagttt
     1921 ctttgtctgg ccatccgggt aacccatgcc ggacgcaaaa tagactactg aaaatttttt
     1981 tgctttgtgg ttgggacttt agccaagggt ataaaagacc accgtccccg aattaccttt
     2041 cctcttcttt tctctctctc cttgtcaact cacacccgaa atcgttaagc atttccttct
     2101 gagtataaga atcattcaaa ggcgcgccat catggtcata gctgtttcct gtgtgaaatt
     2161 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2221 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2281 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4261 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4321 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
