LOCUS       sequence-60455-e        4672 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 4672)
  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 4672)
  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(203)
FEATURES             Location/Qualifiers
     source          1..4672
                     /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(2451..2467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2451..2467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2464..2486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2475..2491
                     /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(2499..2505,2506..2523,2524..2529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2544..2565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2682..2699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2853..3441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2933..2952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3612..4403,4404..4472))
                     /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     4235..4254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4473..4577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4645..4663
                     /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 ctttttcttt gtctggccat ccgggtaacc catgccggac gcaaaataga
     2281 ctactgaaaa tttttttgct ttgtggttgg gactttagcc aagggtataa aagaccaccg
     2341 tccccgaatt acctttcctc ttcttttctc tctctccttg tcaactcaca cccgaaatcg
     2401 ttaagcattt ccttctgagt ataagaatca ttcaaaggcg cgccatcatg gtcatagctg
     2461 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2521 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2581 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2641 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2761 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2821 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2881 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2941 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3001 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3061 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3361 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3421 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3481 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3721 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4621 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
