LOCUS       sequence-319701-        9191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inactivation of Aurli_150841 (crtIBY) in Aurantiochytrium limacinum 
            MYA-1381 with zeocin resistance driven by native GAPDH 
            promoter/termiator system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9191)
  AUTHORS   Rius M, Rest JS, Filloramo GV, Novak Vanclova AMG, Archibald JM, 
            Collier JL
  TITLE     Horizontal Gene Transfer and Fusion Spread Carotenogenesis Among 
            Diverse Heterotrophic Protists.
  JOURNAL   Genome Biol Evol. 2023 Mar 3;15(3):evad029. doi: 
            10.1093/gbe/evad029.
  PUBMED    36805209
REFERENCE   2  (bases 1 to 9191)
  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"; journalName: "Genome Biol
            Evol."; date: "2023-03-3"; volume: "15(3):evad029. doi"; pages: " 
            10.1093/gbe/evad029"
COMMENT     Sequence Label: Aurli_150841_GZG
FEATURES             Location/Qualifiers
     source          1..9191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1803..1819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1803..1819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1816..1838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1827..1843
                     /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(1851..1857,1858..1875,1876..1881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1896..1917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2034..2051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2205..2793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2285..2304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2964..3755,3756..3824))
                     /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     3587..3606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3825..3929)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3997..4015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4053..4075)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4175..4194
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4388..4410
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4402..4419
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4403..4419
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7797..8171
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 gttttctggg gtggcattct ctgcattgtg gcgactgttt ataccacccc ttgggacaac
       61 tacctcgttt atagtacggt ttggaactac ccaccagaac gtatcttatt tgttctgggt
      121 tatgttccta ttgaagaata catgtttttc tcgctggaga cgatgcttgt tgttgcagtt
      181 tggctacatg tattcaaaag tacgactctg caagaggaag ttcagaagcc agctcggggg
      241 gggggtatac tgccactatt gggtctcatg gttgcatggt gtgttggttt ttacatgttg
      301 cagcatgagc agaccttgta catgggccta attttgagct ggggcatgcc cgtgctcata
      361 ctgcagtggg ttctaggcgc ccacgtcctt caaaagcacg ttcgccctgt gacaatagtg
      421 atactatgct cgttctcata tctgtgtata gttgacactt gggcgattca acagggcatc
      481 tggcatctta gttcgcgaaa tttgttacaa cctgtaggca actatactct gcctctagaa
      541 gaggccctgt tctttctggt gacatcaacc atgtgtacat ggggcctcac actggcaatt
      601 gccctctggg gacggcacat tggtatcgca atttctcttc catcgttaca cgctcaacca
      661 aggcgaacgc accacatgtg gcgtcttact catattgtga agcatggagt tattttattc
      721 acagttattg ctcacccatc gatgtatcga tttattccag tttccctagt gacttgctta
      781 cggtttttgg cttggacaga tctagtagct gcccttctcc tgctgtcatc ctgggagatc
      841 cttcaatact ttgggttcgc tttgcttctc tttttcaccc catacccagt cttgactgtt
      901 ctgatctatt tgttgatttt cggcgcggtc ttacctaaac atggagtaga ttctaatagt
      961 ttacctttgt ataaaaaagc ctaactataa accattttcg ttaacatctt gctgaatatg
     1021 gggacacatt tgtaagcatt cttatactca ggaatgactt aaagaaaaac taaatgatga
     1081 agtatcgtaa aacttcaggt ctattcctgc tttcactatc gcgaattttg atttttgctg
     1141 ctcagcttca tttatctcct acaactttct agaccatcca tcattaaaag ataggctgaa
     1201 cttcagaaac tcagttattt ctgttcttgt tgtatgacag gttggtgcaa accttcaagt
     1261 gcagcacaga gccaagtatg aaagtttgag attatttggt ataccatgcg atcttcaaac
     1321 tgtgaggtag cctttttagc aaaagattcc cgcgagtttg gcctacaaaa ttagcgaatt
     1381 ctcgaccata gtattaaagg gccagtacct aggctgctac tgagatatat tataacgaaa
     1441 tgagcattgc tatgcgatac tgaaacacga ctatttataa gtcccttaca aacaaacacc
     1501 agagacacct ttttaaatcc caaggtaagg cgattgattc ttttcaaggt tgagtttttt
     1561 tgttttggtt ttgacacaat acagacagcc gaagctagga gtgatgttca agtacctgca
     1621 tgaaaacctc atctttaaat actttcgtat attattccta gacacttgta gtttcgttcc
     1681 cattcatatc ataatttcga agaaaatcat ttccaaactt agcaaaccgg tcactcatca
     1741 gcattaggcc tttcttctgc actcgagaac tctggagcgc atgcaagctt ggcgtaatca
     1801 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1861 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1921 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1981 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2041 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2101 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2161 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2221 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2281 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2341 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2401 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2461 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2521 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2581 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2641 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2701 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2761 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2821 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2881 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2941 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3001 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3061 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3121 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3181 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3241 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3301 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3361 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3421 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3481 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3541 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3601 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3661 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3721 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3781 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3841 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3901 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3961 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4021 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4081 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4141 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4201 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4261 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4321 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     4381 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtaccgcc
     4441 acccaggatg tctgtcagtg gcgagcgtcg tcgccttcgt cgcgtcctgc cttctctgtt
     4501 ccctctcgcg gagttcgtca cgccctgcct cgcgcaggag gtctccaagc tggtctcgaa
     4561 ttgcttggcg tctcttcaac gtacttcaac gtctcctcgt ttatgtacct tggcttccaa
     4621 tcttgcaatc aatcaagcgc tgtcacgcac cgcttggcgg ccgcgagcct cagcaggtcc
     4681 tcggggtggt cgtcgagcgt caggtttgcc tcgttgtgcg ctgcctgatc tcacgctggg
     4741 ttggtttcgt cgctgagttg attgaatttg atggatttct tgattgattg attgatttct
     4801 ttcttgtttg atgttcgctt gttcgtcgca ggcacgattt cctgaccgcg actacgccca
     4861 cagtggagat gccgttgtgg atctctcccg cagcgagcct ccacacagag tcacacagct
     4921 tcccgcagcc aggtgctccg caatgcagtt catctcttat ttgttgccgc tacttgcacc
     4981 ctgaatagcc ttcgtggtgg ccagcttctt tgcgcctacg ttgcagacag agagtcacgc
     5041 tgacgccttt tctgcgcaag ggtcgaggtg aagcgcagtg ctatcgtcag atggttacct
     5101 aggttcttag aattattaat tcttacttca agttgtttag tgttgttatg tttcaagtta
     5161 ttcttttgtt gtttatttat tgttttattt ttgaaactta ttttcaagtt tgagcaagat
     5221 agcagtcacc tagcaagtaa ctaacgattt atgttactgt tattgtttat gagaaaaaga
     5281 ggggaaaatg taatgtcaaa agcatcattg ggtatttctt gacttgcata atacgacacg
     5341 gagaaaagag taatcgcgat cgaagacgtt tgaggcaaaa ctctaacata ctcgagtcga
     5401 agagagcacc ttcggcatcg ttggtgtcct cgtgaccatg aagtcctcgt gctgtggagt
     5461 gctctgtagc cgcggaagtt gctcatccgg tgacggcatg ctcaccataa gaaggaggcc
     5521 tagccgaaga cttcatgcca cactcccgaa gacctttgca acgaccttgg tccttattct
     5581 tgtggtcctt gcactcgccc tgaaccgtag cacggcgctg aaagtgccca gtgtgagaag
     5641 gtctgcgaca agtattaatg atgctgcttc agaacctgct tgggccgttc aacagaacca
     5701 gaaaattgac ccacaagacc ctcctggaac gcaggataga gcgacttcta gagtaattgt
     5761 cattggagca gggtactcag gactctcagc agcatgtgag ctgcgcagac ttggacatga
     5821 tgtcttggtc ttagagaaga atgactatgt gggtggcaga gctcatcagt ttgaagtcaa
     5881 agctgataat ggacagacct tcaagttcga tgcagggccc agctggtact ggatgcctga
     5941 ggtcttcgat cgattttttg cacgatatgg ccgtaaagtt gaagagtttt acacgctaga
     6001 aaggcttgat ccagcgtaca gaatcattcg ttctgaagaa gaagcaacgc ttgatgtgcc
     6061 gggggcctcg gaggaggcat ttctcagctg ggcacgcaag cttgaaggtg agtctaaagg
     6121 aggcctccta gaccgcatga tggacgaagc taaagaaaag tatgaagagg gtatttttaa
     6181 atggatatgg cacccgatgg tgtccatttg ggagatggtc gatttcaacc ttatgcgtgc
     6241 gcctttccaa tacgacatgt tcaatggcta cctggcacat cttcaaaagt atatttctaa
     6301 ttcagatcta cttacggcgc taaaatggcc tgttatcttc cttggtgcgt caccacactc
     6361 tgcaccagca atgtattccc ttatgacgta tggcggccat gcacttggaa cctactaccc
     6421 tagtggaggg ttaagccggt acccttgatc ttgtgagggc tccactggaa ttttccgaca
     6481 caagaaagtg ccagtggaca aagggggaag tgggcctcga aatagcggta tggttatgtg
     6541 agggatacgg ggcggagttc cggcgttccc tcaggccttc ctctctgcgc ctcaggcaag
     6601 tttctaagaa aagtgctttt ggataataat cttgtatgga aatgtgaagc gcaagtgcgc
     6661 agaacctaga aaatctaaaa caaaagaaaa aagagatcgt cacccgagca gcagaatcaa
     6721 ttcactccaa ggtaagtaag gcgtgctgtg cgggagcgcc cccggccagt cagtgttgga
     6781 gcttgggatg agttgcatct gcgcgaaggg ttggcccatc catctcaagt cctttcctgg
     6841 cggtttcgcc cgcgtcacca gccgcctggt gctcctgatg gatggggact ctggatggtc
     6901 gaaagaggat gtgttgtatc tatctgtctg ataaggtaaa aggggacggc ttctgtactt
     6961 tccttcttcg ctcgctcccg atcgtgtctt ctaggatgcg cgcttttgat gtgctgaaga
     7021 ttccgagcac cgtgtgctat gccgtctccg tcttgcctgt gttggcggct agctagcgct
     7081 gagggttgtg gacggccttg ggaattagga aaagaaagat actcgtgacg ttgaaacgtc
     7141 ctgttctttt tcttttctag tattttctat tccagtcttt actttctgtt cctttcattt
     7201 cacttcaagt acatcgtcac ctctatcgat ctagtgtgag ccgaattgag cctcctgtca
     7261 tgtagcagga ggaactacag aggttccgac tggtacgaac aaacgaatcg gcgaacggag
     7321 agtaggcatg aagttgttgt tgtccaagat caaaaagata gaaacgaata tctttcttgt
     7381 tctgctacct acattgaaac ggacatgaaa gacgacaatt ctagatgaag acactgccaa
     7441 aagggaagaa gcgctcggcc accgcagcaa gaaaggcaag agagagaaga taaagtaaat
     7501 tttcgagaga acaaaagaaa tgaatgcagg aaggaaggaa ggaaggaggg gaggtagtga
     7561 aagaacgcgt gacaagattt gtaatgaaga acatggcatg aaagaacgaa caggggggac
     7621 tgacgatttg agggactgat gtgcgcaatt gaatcttttt catttgcatt gcggctgcgg
     7681 cggcaaacaa aacaaaaata attattcggc attcactttg cttgcgttgt tggacaacat
     7741 aaccataaca gaaacaaaag caaggaaacc tagcgcacac ataaacacca agcaaaatgg
     7801 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     7861 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     7921 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     7981 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     8041 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     8101 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     8161 agcaggactg aggggatcct ctagagtcga catgtaccca ataccacacc ggtagcttct
     8221 cgcggcggct gacaagaaag attgttttta cacatttcga ggcattgatg acccttatcg
     8281 acctatcgtc tcagatcata aaatgcacga gcatgaagta cgcgtgttgt gacttgcttg
     8341 cgtcccacct tctagatggc tttcttcttt tcaagtgatt aaaacaccac agtagatgag
     8401 gactcttagt aagcactgaa gaagcgagta aatagccctc atcccgtttc cctcttttct
     8461 aacacacttt gtttggaatt ctaaaatatc ttttatcatc tcttttcatt cacaaaacta
     8521 gtatttctgc attagaaatc attatcctat ctggcacttt cactctgaca agaacttgcg
     8581 tacatggcgg atcctggcaa tcattttact tgtacagacc caagactttg tgagtcagta
     8641 aatagtaaag aaatgcagaa taatcattag ataattgcaa aaccctgatc ttcaaaatgt
     8701 tatatcacaa gtacctacca agacatttgt atcttctttt tgtcttatgc atttttattt
     8761 cccttagcgc aattaaaaat attaaataac tgacgactct tttcttaagg attcgcggtt
     8821 tgctagttgc actattgaaa agagcacggt attaatttca tagttttact ttgttggcaa
     8881 tctcagcaaa catgggtttt aattttaaaa ttaaataata tttgtgttag actcaaaaga
     8941 tacctataca acccttaggg ctcatcgtca acatttgcaa attaaaacga ttcagctcct
     9001 ttggtatggt catgtctgtt tctgatgcac tgaaaaacct tggacgaacg acacatttta
     9061 ttttcattag gacatctaaa aaacgttgta aagagtatac aataagcata aaaaaggaga
     9121 agaaaaaaaa gaaagacata catggttctg ctcttctagt gcttcaaggc atgcgatttt
     9181 tccgccgagc t
//
