LOCUS       sequence-60444-e        5475 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 5475)
  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 5475)
  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-UAS1B16-TEF(1004)
FEATURES             Location/Qualifiers
     source          1..5475
                     /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(3254..3270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3254..3270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3267..3289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3278..3294
                     /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(3302..3308,3309..3326,3327..3332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3347..3368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3485..3502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3656..4244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3736..3755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4415..5206,5207..5275))
                     /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     5038..5057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5276..5380)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5448..5466
                     /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 aggtaccaag gaagcatgcg
      421 gtacccgaat tcctgaggtg tctcacaagt gccgtgcagt cccgccccca cttgcttctc
      481 tttgtgtgta gtgtacgtac attatcgaga ccgttgttcc cgcccacctc gatccggtct
      541 agactgaggt gtctcacaag tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt
      601 agtgtacgta cattatcgag accgttgttc ccgcccacct cgatccgggg atccctgagg
      661 tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt
      721 acattatcga gaccgttgtt cccgcccacc tcgatccggg tcgacctgag gtgtctcaca
      781 agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg tacattatcg
      841 agaccgttgt tcccgcccac ctcgatccgg gagctcctga ggtgtctcac aagtgccgtg
      901 cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc gagaccgttg
      961 ttcccgccca cctcgatccg gtctagactg aggtgtctca caagtgccgt gcagtcccgc
     1021 ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt gttcccgccc
     1081 acctcgatcc ggggatccct gaggtgtctc acaagtgccg tgcagtcccg cccccacttg
     1141 cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc cacctcgatc
     1201 cgggtcgacc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt gcttctcttt
     1261 gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat ccgggcatgc
     1321 ggtacccgaa ttcctgaggt gtctcacaag tgccgtgcag tcccgccccc acttgcttct
     1381 ctttgtgtgt agtgtacgta cattatcgag accgttgttc ccgcccacct cgatccggtc
     1441 tagactgagg tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg
     1501 tagtgtacgt acattatcga gaccgttgtt cccgcccacc tcgatccggg gatccctgag
     1561 gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg
     1621 tacattatcg agaccgttgt tcccgcccac ctcgatccgg gtcgacctga ggtgtctcac
     1681 aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc
     1741 gagaccgttg ttcccgccca cctcgatccg ggagctcctg aggtgtctca caagtgccgt
     1801 gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt
     1861 gttcccgccc acctcgatcc ggtctagact gaggtgtctc acaagtgccg tgcagtcccg
     1921 cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc
     1981 cacctcgatc cggggatccc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt
     2041 gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat
     2101 ccgggtcgac ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt
     2161 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccgggcatg
     2221 cctgcagaag ctttctgtaa aaagtctcta caagtagtct tgtaaaaaca ttccccacag
     2281 taaccatcta tattgcaaaa caatacgact gcaattgggg attgttgctc gtatcgcacc
     2341 cgtaccgtgc caccgtaaca ccccccagac accaatcact acgaccgatc caactccacg
     2401 tttctgctac ccagtctgct ggccggcttg gctttgcgtt tcgctcccac actacaccgt
     2461 aataccacgt cactctcatt gcaggttacc ctgcccgtag tcgctcgatc cacctcctcc
     2521 ttctctcgtg tgtgcagcaa agaggcagag atggagcccg tatggtgagc cgtagagtgg
     2581 agcgaggggc gatcttacaa cggctgtcgg cggatataaa cggtattttc acaattgcac
     2641 cccagccaga ccgatagccg gtcgcaatcc gccacccaca accgtctacc tcccacagaa
     2701 ccccgtcact tccacccttt tccaccagat catatgtccc aacttgccaa attaaaaccg
     2761 tgcgaatttt caaaataaac tttggcaaag aggctgcaaa ggaggggctg gtgagggcgt
     2821 ctggaagtcg accagagacc gggttggcgg cgcatttgtg tcccaaaaaa cagccccaat
     2881 tgccccaatt gaccccaaat tgacccagta gcgggcccaa ccccggcgag agcccccttc
     2941 tccccacata tcaaacctcc cccggttccc acacttgccg ttaagggcgt agggtactgc
     3001 agtctggaat ctacgcttgt tcagactttg tactagtttc tttgtctggc catccgggta
     3061 acccatgccg gacgcaaaat agactactga aaattttttt gctttgtggt tgggacttta
     3121 gccaagggta taaaagacca ccgtccccga attacctttc ctcttctttt ctctctctcc
     3181 ttgtcaactc acacccgaaa tcgttaagca tttccttctg agtataagaa tcattcaaag
     3241 gcgcgccatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3301 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3361 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3421 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3481 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3541 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3661 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4081 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4321 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4381 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4441 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4501 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4561 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4621 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4681 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4741 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4801 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4861 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4921 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4981 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5041 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5101 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5161 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5221 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5281 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5341 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5401 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5461 cgaggccctt tcgtc
//
