LOCUS       sequence-60458-e        3822 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 3822)
  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 3822)
  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(272)
FEATURES             Location/Qualifiers
     source          1..3822
                     /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(1601..1617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1601..1617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1614..1636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1625..1641
                     /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(1649..1655,1656..1673,1674..1679))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1694..1715
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1832..1849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2003..2591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2083..2102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2762..3553,3554..3622))
                     /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     3385..3404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3623..3727)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3795..3813
                     /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 gcttcccaca
     1321 cttgccgtta agggcgtagg gtactgcagt ctggaatcta cgcttgttca gactttgtac
     1381 tagtttcttt gtctggccat ccgggtaacc catgccggac gcaaaataga ctactgaaaa
     1441 tttttttgct ttgtggttgg gactttagcc aagggtataa aagaccaccg tccccgaatt
     1501 acctttcctc ttcttttctc tctctccttg tcaactcaca cccgaaatcg ttaagcattt
     1561 ccttctgagt ataagaatca ttcaaaggcg cgccatcatg gtcatagctg tttcctgtgt
     1621 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1681 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1741 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1801 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2401 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2821 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3601 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3721 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     3781 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
