LOCUS       sequence-60310-e        3901 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 3901)
  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 3901)
  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-UAS1B10-Leum
FEATURES             Location/Qualifiers
     source          1..3901
                     /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(1680..1696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1680..1696)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1693..1715)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1704..1720
                     /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(1728..1734,1735..1752,1753..1758))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1773..1794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1911..1928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2082..2670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2162..2181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2841..3632,3633..3701))
                     /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     3464..3483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3702..3806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3874..3892
                     /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 ggggatccct
     1321 gaggtgtctc acaagtgccg tgcagtcccg cccccacttg cttctctttg tgtgtagtgt
     1381 acgtacatta tcgagaccgt tgttcccgcc cacctcgatc cgggtcgacc tgaggtgtct
     1441 cacaagtgcc gtgcagtccc gcccccactt gcttctcttt gtgtgtagtg tacgtacatt
     1501 atcgagaccg ttgttcccgc ccacctcgat ccgggcatgc actgatcacg ggcaaaagtg
     1561 cgtatatata caagagcgtt tgccagccac agattttcac tccacacacc acatcacaca
     1621 tacaaccaca cacatccaca atggaacccg aaactaaaag cttcaggcgc gccatcatgg
     1681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2521 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3841 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3901 c
//
