LOCUS       sequence-60445-e        4556 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 4556)
  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 4556)
  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(1004)
FEATURES             Location/Qualifiers
     source          1..4556
                     /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(2335..2351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2335..2351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2348..2370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2359..2375
                     /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(2383..2389,2390..2407,2408..2413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2428..2449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2566..2583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2737..3325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2817..2836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3496..4287,4288..4356))
                     /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     4119..4138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4357..4461)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4529..4547
                     /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 gctttctgta
     1321 aaaagtctct acaagtagtc ttgtaaaaac attccccaca gtaaccatct atattgcaaa
     1381 acaatacgac tgcaattggg gattgttgct cgtatcgcac ccgtaccgtg ccaccgtaac
     1441 accccccaga caccaatcac tacgaccgat ccaactccac gtttctgcta cccagtctgc
     1501 tggccggctt ggctttgcgt ttcgctccca cactacaccg taataccacg tcactctcat
     1561 tgcaggttac cctgcccgta gtcgctcgat ccacctcctc cttctctcgt gtgtgcagca
     1621 aagaggcaga gatggagccc gtatggtgag ccgtagagtg gagcgagggg cgatcttaca
     1681 acggctgtcg gcggatataa acggtatttt cacaattgca ccccagccag accgatagcc
     1741 ggtcgcaatc cgccacccac aaccgtctac ctcccacaga accccgtcac ttccaccctt
     1801 ttccaccaga tcatatgtcc caacttgcca aattaaaacc gtgcgaattt tcaaaataaa
     1861 ctttggcaaa gaggctgcaa aggaggggct ggtgagggcg tctggaagtc gaccagagac
     1921 cgggttggcg gcgcatttgt gtcccaaaaa acagccccaa ttgccccaat tgaccccaaa
     1981 ttgacccagt agcgggccca accccggcga gagccccctt ctccccacat atcaaacctc
     2041 ccccggttcc cacacttgcc gttaagggcg tagggtactg cagtctggaa tctacgcttg
     2101 ttcagacttt gtactagttt ctttgtctgg ccatccgggt aacccatgcc ggacgcaaaa
     2161 tagactactg aaaatttttt tgctttgtgg ttgggacttt agccaagggt ataaaagacc
     2221 accgtccccg aattaccttt cctcttcttt tctctctctc cttgtcaact cacacccgaa
     2281 atcgttaagc atttccttct gagtataaga atcattcaaa ggcgcgccat catggtcata
     2341 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2401 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2461 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2521 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2581 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2641 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2701 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2761 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2821 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2881 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2941 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3001 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3061 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3121 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3181 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3781 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4081 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4501 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
