LOCUS       sequence-437868-        6871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HisG-URA3-HisG recycling type marker in pTAs for Yarrowia 
            lipolytica.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6871)
  AUTHORS   Gao S, Tong Y, Wen Z, Zhu L, Ge M, Chen D, Jiang Y, Yang S
  TITLE     Multiplex gene editing of the Yarrowia lipolytica genome using the 
            CRISPR-Cas9 system.
  JOURNAL   J Ind Microbiol Biotechnol. 2016 Aug;43(8):1085-93. doi: 
            10.1007/s10295-016-1789-8. Epub 2016 Jun 27.
  PUBMED    27349768
REFERENCE   2  (bases 1 to 6871)
  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: 
            "10.1007/s10295-016-1789-8"; journalName: "J Ind Microbiol 
            Biotechnol"; date: "2016-08"; volume: "43"; issue: "8"; pages: 
            "1085-93"
COMMENT     Sequence Label: pTA-HUH
FEATURES             Location/Qualifiers
     source          1..6871
                     /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(4650..4666)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4650..4666)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4663..4685)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4674..4690
                     /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(4698..4704,4705..4722,4723..4728))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4881..4898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5052..5640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5132..5151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5811..6602,6603..6671))
                     /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     6434..6453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6672..6776)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6844..6862
                     /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 ccagtgccaa gcttggggct gttccagtca
      421 atcagggtat tgaagctcat ggtctttact ccatcacagg gttccgcctt atccggccta
      481 cagaacccaa aatatcaacg cattacgtag gcctgataag cgcagcgcca tcaggcgtca
      541 gatcactcca tcatcttctc gatcggcagt accagaatcg agctggcgcc aagcgctttc
      601 agtttctcca tggtttccca gaacaacgtt tcgctgctga ccatgtgcat cgccacgcgc
      661 tgttgctcgc ctgccagcgg cagaattgtc ggcctttcgg cgcctggcag cagggcgata
      721 acctcttcca ggcgttcact tggcgcgtgc atcatgatgt atttcgattc gcgcgcctga
      781 atcacgccct gaatacgggt cagcaattta tcgatcagct cttgcttgct ctgtgccatc
      841 tcaccgtcgc gctgaatcag acaggcttta gagcggtaga taacttcgac ttcacgcagg
      901 ccgttagctt caagcgtcgc gccggtagag accaaatcgc agatagcgtc ggccagcccc
      961 gcgcgcggcg cgacttcgac agaaccattt aacagacacg atttaaaaga gacgcctttc
     1021 tggtcgaggt agcgtttgag gaggtgcgga tatgaggtag cgatacgttt accgtccagc
     1081 gcggccgggc cgtcccaggc ttcgtcaacc ggtgttgcca gcgataaacg gcagccgccg
     1141 aagtcaagac ggcgcagggt taaatagcgt ggatcttcgc cctgtgcgcg gcggttgagt
     1201 agctcttctt ccagcacgtt ttcgccgata ataccgagat cgaccacgcc atccattacc
     1261 agacccggaa tgtcatcatc acgcacgcgc aggatatcaa tcggcatgtt ttccgccatc
     1321 gcaatcaggc gctgagtgtg taaattaatt tttatgccgc agcgggccag caattctcgt
     1381 gaatcatcgc ttaaacggcc tgatttctga atagctatgc gtaagcgggt gttgtctaac
     1441 attctgcgtt cctctttatc ctgtctgaac cggtctgtat cgcgcgccaa aaaaaaagcc
     1501 cccggaagat gatcttccgg gggctttctc atgcgttcat gcaccactgg aagatccggg
     1561 atcccgggtg tagtggtagt gcagtggtgg tattgtgact ggggatgtag ttgagaataa
     1621 gtcatacaca agtcagcttt cttcaagcct catataagta taagtagttc aatcgtatta
     1681 gctacttgta cccagcatct ccgtatcgag aaacacaaca acatgcccca ttggacagat
     1741 catgcggata cacaggttgt gcagtatcat acatactcga tcagacaggt cgtctgacca
     1801 tcatacaagc tgaacaagcg ctccatactt gcacgctctc tatatacaca gttaaattac
     1861 atatccatag tctaacctct aacagttaat cttctggtaa gcctcccagc cagccttctg
     1921 gtatcgcttg gcctcctcaa taggatctcg gttctggccg tacagacctc ggccgacaat
     1981 tatgatatcc gttccggtag acatgacatc ctcaacagtt cggtactgct gtccgagagc
     2041 gtctcccttg tcgtcaagac ccaccccggg ggtcagaata agccagtcct cagagtcgcc
     2101 cttaggtcgg ttctgggcaa tgaagccaac cacaaactcg gggtcggatc gggcaagctc
     2161 aatggtctgc ttggagtact cgccagtggc cagagagccc ttgcaagaca gctcggccag
     2221 catgagcaga cctctggcca gcttctcgtt gggagagggg accaggaact ccttgtactg
     2281 ggagttctcg tagtcagaga cgtcctcctt cttctgttca gagacagttt cctcggcacc
     2341 agctcgcagg ccagcaatga ttccggttcc gggtacaccg tgggcgttgg tgatatcgga
     2401 ccactcggcg attcggtaga caccgttctt gtactggtgc ttgacagtgt tgccaatatc
     2461 tgcgaacttt ctgtcctcga acaggaagaa accgtgctta agagcaagtt ccttgagggg
     2521 gagcacagtg ccggcgtagg tgaagtcgtc aatgatgtcg atatgggtct tgatcatgca
     2581 cacataaggt ccgaccttat cggcaagctc aatgagctcc ttggtggtgg taacatccag
     2641 agaagcacac aggttggttt tcttggctgc cacgagcttg agcactcgag cggcaaaggc
     2701 ggacttgtgg acgttagctc gagcttcgta ggagggcatt ttggtggtga agaggagact
     2761 gaaataaatt tagtctgcag aactttttat cggaacctta tctggggcag tgaagtatat
     2821 gttatggtaa tagttacgag ttagttgaac ttatagatag actggactat acggctatcg
     2881 gtccaaatta gaaagaacgt caatggctct ctgggcgtcg cctttgccga caaaaatgtg
     2941 atcatgatga aagccagcaa tgacgttgca gctgatattg ttgtcggcca accgcgccga
     3001 aaacgcagct gtcagaccca cagcctccaa cgaagaatgt atcgtcaaag tgatccaagc
     3061 acactcatag ttggagtcgt actccaaagg cggcaatgac gagtcagaca gatactcgtc
     3121 gaccttttcc ttgggaacca ccaccgtcag cccttctgac tcacgtattg tagccaccga
     3181 cacaggcaac agtccgtgga tagcagaata tgtcttgtcg gtccatttct caccaacttt
     3241 aggcgtcaag tgaatgttgc agaagaagta tgtgccttca ttgagaatcg gtgttgctga
     3301 tttcaataaa gtcttgagat cagtttggcc agtcatgttg tggggggtaa ttggattgag
     3361 ttatcgccta cagtctgtac aggtatactc gctgcccact ttatactttt tgattccgct
     3421 gcacttgaag caatgtcgtt taccaaaagt gagaatgctc cacagaacac accggggtac
     3481 cccgctgttc cagtcaatca gggtattgaa gctcatggtc tttactccat cacagggttc
     3541 cgccttatcc ggcctacaga acccaaaata tcaacgcatt acgtaggcct gataagcgca
     3601 gcgccatcag gcgtcagatc actccatcat cttctcgatc ggcagtacca gaatcgagct
     3661 ggcgccaagc gctttcagtt tctccatggt ttcccagaac aacgtttcgc tgctgaccat
     3721 gtgcatcgcc acgcgctgtt gctcgcctgc cagcggcaga attgtcggcc tttcggcgcc
     3781 tggcagcagg gcgataacct cttccaggcg ttcacttggc gcgtgcatca tgatgtattt
     3841 cgattcgcgc gcctgaatca cgccctgaat acgggtcagc aatttatcga tcagctcttg
     3901 cttgctctgt gccatctcac cgtcgcgctg aatcagacag gctttagagc ggtagataac
     3961 ttcgacttca cgcaggccgt tagcttcaag cgtcgcgccg gtagagacca aatcgcagat
     4021 agcgtcggcc agccccgcgc gcggcgcgac ttcgacagaa ccatttaaca gacacgattt
     4081 aaaagagacg cctttctggt cgaggtagcg tttgaggagg tgcggatatg aggtagcgat
     4141 acgtttaccg tccagcgcgg ccgggccgtc ccaggcttcg tcaaccggtg ttgccagcga
     4201 taaacggcag ccgccgaagt caagacggcg cagggttaaa tagcgtggat cttcgccctg
     4261 tgcgcggcgg ttgagtagct cttcttccag cacgttttcg ccgataatac cgagatcgac
     4321 cacgccatcc attaccagac ccggaatgtc atcatcacgc acgcgcagga tatcaatcgg
     4381 catgttttcc gccatcgcaa tcaggcgctg agtgtgtaaa ttaattttta tgccgcagcg
     4441 ggccagcaat tctcgtgaat catcgcttaa acggcctgat ttctgaatag ctatgcgtaa
     4501 gcgggtgttg tctaacattc tgcgttcctc tttatcctgt ctgaaccggt ctgtatcgcg
     4561 cgccaaaaaa aaagcccccg gaagatgatc ttccgggggc tttctcatgc gttcatgcac
     4621 cactggaaga tccggaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5461 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5881 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5941 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6001 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6061 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6661 tgaatactca tactcttcct ttttcaatct tattgaagca tttatcaggg ttattgtctc
     6721 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6781 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6841 aaaaataggc gtatcacgag gccctttcgt c
//
