LOCUS       sequence-60450-e        4975 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 4975)
  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 4975)
  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(504)
FEATURES             Location/Qualifiers
     source          1..4975
                     /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(2754..2770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2754..2770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2767..2789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2778..2794
                     /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(2802..2808,2809..2826,2827..2832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2847..2868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2985..3002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3156..3744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3236..3255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3915..4706,4707..4775))
                     /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     4538..4557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4776..4880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4948..4966
                     /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 cttatgtccc aacttgccaa attaaaaccg tgcgaatttt caaaataaac
     2281 tttggcaaag aggctgcaaa ggaggggctg gtgagggcgt ctggaagtcg accagagacc
     2341 gggttggcgg cgcatttgtg tcccaaaaaa cagccccaat tgccccaatt gaccccaaat
     2401 tgacccagta gcgggcccaa ccccggcgag agcccccttc tccccacata tcaaacctcc
     2461 cccggttccc acacttgccg ttaagggcgt agggtactgc agtctggaat ctacgcttgt
     2521 tcagactttg tactagtttc tttgtctggc catccgggta acccatgccg gacgcaaaat
     2581 agactactga aaattttttt gctttgtggt tgggacttta gccaagggta taaaagacca
     2641 ccgtccccga attacctttc ctcttctttt ctctctctcc ttgtcaactc acacccgaaa
     2701 tcgttaagca tttccttctg agtataagaa tcattcaaag gcgcgccatc atggtcatag
     2761 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2821 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2881 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4921 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
