LOCUS       sequence-460292-        9242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NIT-9E domain in N. crassa at the his-3 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9242)
  TITLE     Convergent Evolution Links Molybdenum Insertase Domains with 
            Organism-Specific Sequences.
REFERENCE   2  (bases 1 to 9242)
  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"
COMMENT     Sequence Label: pCCG_nit9E
FEATURES             Location/Qualifiers
     source          1..9242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     217..239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(277..295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        363..467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(468..536,537..1328)
                     /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     complement(686..705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1499..2087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1988..2007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2241..2258
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2638..2649
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4007..4025
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6525..6541)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6526..6542)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(join(9241..9242,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(9242,1..17))
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc
       61 gttctaacga caatatgtcc atatggtgca ctctcagtac aatctgctct gatgccgcat
      121 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      181 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      241 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      301 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      541 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      601 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      661 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      781 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      841 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      901 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1021 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1081 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1201 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1261 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1321 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1381 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1441 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1561 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1621 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1681 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1741 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1801 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1861 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1921 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1981 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2041 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2101 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2161 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2221 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2281 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggttaacc tggcttatcg
     2341 aaattaatac gactcactat agggagaccg gcagatccaa tgcggatgga ttcgctcact
     2401 cggtgccgcg gcggtgcgaa gttcctgccg tggattgggg gattgcacag acggtggagt
     2461 gcttgctaca gctaggtacc cagttgcaca ccaaacggtg cccctgtgcc cctgcgtcta
     2521 ggtgtgtcct gtgagcaact gcgcccgcgc gcggcaccgg gaagctgcgg gaacctcaat
     2581 ggatgattgg cgccagatcc ccggcatcac ctcagcacat gggcagcccg agaaggaatc
     2641 gagggtagaa gatattccag ggtttccgct agcacaactt ccaagcgcct ctaccgctac
     2701 tccacacacc cacgctgaat ggcacaacca ctgaggccta gcggtcccag ctccccgtaa
     2761 ccacgcagcc ccagcgaccg aggtcatcat gacactatga cgacaatctg gcgggagact
     2821 tgggaaaatc gatgacgggc acctaccttg ggaacaatag agcaagtcca gtcaacgccc
     2881 tagggaaggg ggaaaaaaaa aaaagagccc attctcacaa ggagcaacac tgtgctgcta
     2941 tcatagtaca ctacaagcaa agtcgggatg ttgctagaat ccgtcggcta cgtcggtaga
     3001 tacctcccat aggcagtaag tctaggtatc caatttatag ttccgcgtat ttcttcgtcg
     3061 aacggggcac agcaaagcct aacagccgca gtgcctcgca accaacacca agcagtaatc
     3121 tcatccatcc atacagatcc tcccacaata tctttacatg cgcacttgcg ctgtgttagc
     3181 agtttaggtt ccactaccaa ggctgacacg agcagcactg acagcaaact gctgcttggg
     3241 gccaagaaat tggcagtttg ctgagcacct acatatcagt acacatcaac cctggtttaa
     3301 ttgttagcct ggtactgtta accgtcatga acgaggacgt ggaggaagaa ccacgctaac
     3361 aattgaacca gggttgatgt atacctctct taccatattt gcttgcttcc acgtccctat
     3421 cctcaagcga ttttgttctt gactttttgt atttcaacgc aaaggccatt caggtttctc
     3481 atgttcaacc ctttggatgg cgatgccgtt cgtcccagga cctcgtgacc tttgaaagac
     3541 gagaaatata tcccaaagca acttctgctt atggacaagc aactgcatgg cggcccacaa
     3601 ctcatatctc gcgtaatcat ccaaacgcta ggcccgagta ctctactgtt ctcgagacac
     3661 ctaccataat atgtacccaa tcagacatga caacagtcaa ccaaacatac cctgatccgg
     3721 ccaaaccctg acatccctag atgatggtta ggcacacaaa tggaaaacgg gactcgttgg
     3781 ttaggtggga acgcttgtat tgcagccagc caaccgactc tcctcgtccc tctcgagtcc
     3841 cgttattgcc gtttgaccca gccgcccagc cgccaaacag ctgagggagc caatggtcta
     3901 atacaccgtc caacggcccc agtgatgttt caaacttgat tgacagcgaa cgaaacccct
     3961 gaaacccgtt atagatatgt atagatttga cggggtagct tggccctaat acgactcact
     4021 atagggcgaa ttggagctcc accgcggtgg cggccgctag aaggagcagt ccatctgcgt
     4081 gaatcacgag agaatcagct actttgaatc gatggatgca gctaccagaa gtcactcagt
     4141 tcgttcaaag ccacatcact gggcacttcc attgggacag gcattgatcg gacgagaccg
     4201 acttctggcc gctttcaaca gccacattat atccatgtca cggctacgcg cggccttcgg
     4261 taacagaaaa gcacacagac agcgattgtg acatggattc gggcaaagca ttggtggtcg
     4321 caccaggtca tcctgagtgt gcagtggctg ctattcagat ttcatctaac tgcgggagag
     4381 gggttcaaag gggcgtgacg tcacagacaa cgggtgaagg acgaagattg cctcacttct
     4441 ttgctagcaa tttgcctgca aagaaggcgc acatgtcaag caaacaactg ggaaaccact
     4501 attgaatacc cacaatgcaa agctcggaag gtacgtcttg attgcagtgt gtcgagtgtc
     4561 aaaaaagaag caagtgttca tgcaagccaa aattggcacc tcctccactt ctccgagtgc
     4621 cccacccgaa cctccaggcg agatggccgg aacataccat ccgcgttggg attatgacgt
     4681 atctccttct tcttcacatg attccatccc gttgttgctt gtttgcgagc tgtgacggga
     4741 gatcgtagat gccacttagg gccaggcagg cagtgcaggc agccaggaac acaagcttcc
     4801 aacttggtca tctcgattgc cgattaaggg aaccaaatgc ctatataaga ctgtcctccc
     4861 acctccccaa taccattctt ttcttcttcc atcatcagcc aacaaagcaa tcacatcttc
     4921 actacttcaa atcaacacaa cactcaaacc actttcacaa cccctcacat caaccaaaat
     4981 gctctctgtc tccgaagcgc tctcccaaat agcgcaacac acgccgatgc cgcaaatcgt
     5041 cacctccaaa gtcgatagct cgctcatcgg cgccgttctg gcgcaggacg tttcggcgcg
     5101 cgaaaatgtg cccgcttttc gcgctagcat cgtcgacggc tacgccgtag tcgtgcccca
     5161 agacgggaac atgcgcggcg tgttccccgt cgtgtcggtt tcgcatgccg ctcctggaga
     5221 tggggaaaaa atgccgacgc tcaaagaggg cgagattgcc cggatcacca cgggtgcacc
     5281 cctgcctccg ggtgcgacat cggttattat ggtggaggat acggtgttgg tgtcgacgac
     5341 gtcatccaag acggaggatg cggaggatgg aaagggagtg gaggaggaga aggaggtgga
     5401 gattttagca gctgataaag ttcgcgaagg cgaaaatgtt agggaggtgg gaagtgatat
     5461 caagcagggg caactgattc taggcaaggg agatcaggtg tctgctgtgg ggggtgagat
     5521 tggtctgttg gccgccgtgg gtgtgggtga ggtgcaggtg taccgacgtc ctgttgttgg
     5581 cgttctatct acgggtgatg agattgtgga ccatgatcgc ccgggagact tgcggctggg
     5641 cgaggtgcgc gatacgaacc ggccaactct catttcggcg gcgagagatt gggggtatga
     5701 ggtgattgat cttggagtgg cgcgggacaa ggccgggact ttggaggaga cgctgcggca
     5761 ggcattgagg caggttgacg tggtgatcac gaccggcgga gtgtccatgg gtgagctgga
     5821 tctcctcaag ccgacgatag agcgttcgct gggcggaaca attcgctttg gtcgggtggc
     5881 aatgaagccc ggcaagccta ccacgtttgc taccgttccg gtaaaggata acgacggaga
     5941 gagagtttcc aaggtaatct tctcgctacc aggtaacccg gcatcggctt tggttacgtt
     6001 tcacttgttt gtactgccgt ctctgcacca aatgtcgggc gtctcaccac ctggcttgcc
     6061 aaaggtgcct gtgacactgg gccatgattt ctccctcgat agatctagac ccgagtacca
     6121 ccgggccatt gtctccgtta gcagggacgg agtgctcacc gcggtcagca caggaggaca
     6181 gagaagctcg cgggtaggaa gtatgaaggg ggccaacgcg cttctctgca tgccttgcgg
     6241 accagagccg ttgcgcaagg gtgccaaggt ggaagcgttg cttatgggcg gtttacgcag
     6301 cgatgtgatt tagtgatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     6361 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     6421 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     6481 aggggcaagt tgggaagaca acctgaattc gatatcaagc ttatcgatac cgtcgacctc
     6541 gagggggggc ccggtaccag cttttgttcc ctttagtgag ggttaatagg gcgtgcacgg
     6601 ctatggggtc tgaatgctaa aagacaccat ttcccacact ccctctgatt ttttttatgg
     6661 ttttttttta tcttcatttt tcgccgtctt ttattcggtt gtttgggagg ttcacatgca
     6721 acgtgcaaga ctgcaagtat attgagatgg gcgcttcatg caatgcagca tcactgcagc
     6781 gacattgtga catccacgtt tcgatcgtct gtgcttcccc gttcatgtac gcatgatctg
     6841 aaaaaaaaaa aaaaaaaaaa gttgatgtaa taatgaaaat aaacacccag ggacgagcac
     6901 cccttctgga atagggagaa aaactaaagg gacgtttggg tctctacact taccaacaga
     6961 ttcatgtact gatcttcagc ctgtacagaa cagtacaatg tccaaaactc gttatcatta
     7021 ctctatgtgc ccaaaatgac ccagatatac atcaacccac tacatccttt tgttcttgtt
     7081 tttttttttt tttttgctac ttcgaaaaga atttccgttt agttgctctt gctcatgtgc
     7141 tcaagacgga tgctgaccgc ccttctgtga gcctcgagct cctcaacctt agccaactgc
     7201 atgacagcct ggccgacgtt tttgagaccc tcggcagtca agttggagct ggtaatgtgc
     7261 ttgacgaacg aggcgagatt gacgccagag tactgcttgg caaagccata ggtagctatg
     7321 tgtcttgacg ttagcagaag aatctgtgaa gcggggctat ttatgatatg taacttacgc
     7381 agcgagtggt taacaccagc agagtaatcg ccaacggact caggagtcca ggcgccaatg
     7441 aagacactac cggcgttcat gacaagatcg acagccttct cggcctcctt gatctggagg
     7501 atcaagtgct caggagcgta cttgttgctg agctccatgg cctcctcgac ggtcttgacc
     7561 tgcacggtga tcgagtgggc gatggagcca cggacaatct ggacgcgagg aagctccgta
     7621 gcctgacggt gaacctcgtc ctcaatagcc tgaagatgct cctcgtcgag gtcaatagcg
     7681 atcaggatga cctgactgtc aacgccgtgc tcagcctggg acaggagatc cgaggcaacg
     7741 aacgcggggt tggcgtcctt gtcagcgatg accagcacct cggacgggcc agcgggcatg
     7801 tcaataccaa cggcagcgtt ggtgtcgttg ctgacgaaca tcttggcagc agtgacgaac
     7861 tggttaccgg ggccgagaat cttgtcgacc ttggtgatgc tctcggtgcc gtaggccatg
     7921 gcagctacgg cctgggcacc gccggcaagc acgatggact cggcgccaac cttgtgagcg
     7981 acgtagacaa tctcgggagt gatggttccg tcggcgcggg gaggagaggc gaacacaatc
     8041 ttgttgcagc cggcgaccat ggcgggaaca cccagcataa gggcagtgct ggggagaacg
     8101 gcggtaccgc cggggatgta gcagccgacg gcctcgatgg gacgagagaa acggctgcag
     8161 acaacaccgg gcatggtctc gacctggagg ggcttctcct ccttctgggc ggcgtggaac
     8221 ttgcggatgt tctcgaagga cacgtcgatg gcagcaatgg tctcctcagg gagctgcata
     8281 agctccttgg ggaagggcgc cttcaggacg gggctagtaa gagaggtagc cttctcgaac
     8341 ttgtgagtgt acgacagaac agccttgtcg ccgttcttgc ggacgtcctc gatgatgggg
     8401 acaatgatct tgtagatggc atcggacgac ttttgcgcag gacgcttgag agcagcatcg
     8461 agctcctcgg tagagacctt ggaggcgtcg aaacgtctca tggtgatctt ctcaggggtg
     8521 gtctcctggg cggcctcctt ggtgacaggg gccgaagtgg cagcgagagc ggacgccgca
     8581 ggcttgatgc cctccttctc agcccactta cccttagcat ctccagtcct gcgcttgacc
     8641 ttccagctct tggcgtcaag gctcctttcg atatcggcaa gagtaacgcc ggcagcaacg
     8701 gccctggtaa gagcaaagta gaagagatcg gcagcctcaa aggcgatttc ctggggggtc
     8761 tgagcggtgc agagctcctc agcctcctcc atgatcttgg cccggactag cttctcatcg
     8821 gagaagagac gggcagtgta ggagccctcg ggggcagact gtttcctcga aatcaaagtc
     8881 tgctcgagct tgggaaggcc tttgagctga ccaaagcagc cggactgatc gaggtggcag
     8941 aaacgaccct tctgcttcac gacaaacttg agagcatcgt tatcgcagtc aagcgagatg
     9001 cggacgagct cctgagtgtc tccggaagta gcacccttgt accagagacc gcgcttccgg
     9061 ctctgataga caccagtctg tgtcctgagg gcctcgttca cactctcggc actgctgtag
     9121 accagaccca gagcagtgtc gtgctcatca acgacaacgg tggggagaag accatcggga
     9181 cggtcagact tccagacgct cgagaggatg gtggagatgg caagcttcag ctgctcgagt
     9241 tc
//
