LOCUS       sequence-459450-        9931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NIT-9 with reversed domains in N. crassa at the his-3 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9931)
  TITLE     Convergent Evolution Links Molybdenum Insertase Domains with 
            Organism-Specific Sequences.
REFERENCE   2  (bases 1 to 9931)
  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_nit9fl_reverse
FEATURES             Location/Qualifiers
     source          1..9931
                     /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     4979..4995
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4979..4995
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7212..7228)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7213..7229)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(join(9930..9931,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(9931,1..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 caaccaaatc
     4981 tagaactagt ggatccccgg gttaattaac atgctctctg tctccgaagc gctctcccaa
     5041 atagcgcaac acacgccgat gccgcaaatc gtcacctcca aagtcgatag ctcgctcatc
     5101 ggcgccgttc tggcgcagga cgtttcggcg cgcgaaaatg tgcccgcttt tcgcgctagc
     5161 atcgtcgacg gctacgccgt agtcgtgccc caagacggga acatgcgcgg cgtgttcccc
     5221 gtcgtgtcgg tttcgcatgc cgctcctgga gatggggaaa aaatgccgac gctcaaagag
     5281 ggcgagattg cccggatcac cacgggtgca cccctgcctc cgggtgcgac atcggttatt
     5341 atggtggagg atacggtgtt ggtgtcgacg acgtcatcca agacggagga tgcggaggat
     5401 ggaaagggag tggaggagga gaaggaggtg gagattttag cagctgataa agttcgcgaa
     5461 ggcgaaaatg ttagggaggt gggaagtgat atcaagcagg ggcaactgat tctaggcaag
     5521 ggagatcagg tgtctgctgt ggggggtgag attggtctgt tggccgccgt gggtgtgggt
     5581 gaggtgcagg tgtaccgacg tcctgttgtt ggcgttctat ctacgggtga tgagattgtg
     5641 gaccatgatc gcccgggaga cttgcggctg ggcgaggtgc gcgatacgaa ccggccaact
     5701 ctcatttcgg cggcgagaga ttgggggtat gaggtgattg atcttggagt ggcgcgggac
     5761 aaggccggga ctttggagga gacgctgcgg caggcattga ggcaggttga cgtggtgatc
     5821 acgaccggcg gagtgtccat gggtgagctg gatctcctca agccgacgat agagcgttcg
     5881 ctgggcggaa caattcgctt tggtcgggtg gcaatgaagc ccggcaagcc taccacgttt
     5941 gctaccgttc cggtaaagga taacgacgga gagagagttt ccaaggtaat cttctcgcta
     6001 ccaggtaacc cggcatcggc tttggttacg tttcacttgt ttgtactgcc gtctctgcac
     6061 caaatgtcgg gcgtctcacc acctggcttg ccaaaggtgc ctgtgacact gggccatgat
     6121 ttctccctcg atagatctag acccgagtac caccgggcca ttgtctccgt tagcagggac
     6181 ggagtgctca ccgcggtcag cacaggagga cagagaagct cgcgggtagg aagtatgaag
     6241 ggggccaacg cgcttctctg catgccttgc ggaccagagc cgttgcgcaa gggtgccaag
     6301 gtggaagcgt tgcttatggg cggtttacgc agcgatgtga ttgctttaca taagggggga
     6361 gtgaagaaat tggagcagga ggcgggagta gaaggaggtg gtggacgtgg acatggtcac
     6421 ggacatggac acggacatgg acacaagcat cacgaccatg gcgagcaccg aggaggagga
     6481 gggggaggct gtggcgccca tgggcatggc cataaccacg gtcacggccc aggccacggc
     6541 cacgccggtc tagtccgaca caccatcccc tctgaaaacc ccaagtccaa cgatcccagc
     6601 ttaggtccca gccgccgcca ccgcgagtct ccttacccga tgtcctcaag tactactcca
     6661 gcagcgctca aagcagccat ccttatcgtt tcaacgacag cagctcaaga ccccgcaacg
     6721 gactcggccg acgtcactct ccgccaggtc ttcgatgaac aacaagccca atgggaggtc
     6781 catgaatcca aaatcgttcc cgacgatgtc gtgcaaatcc agaagcaaat catccactgg
     6841 acaaaccaag ccaccgccgc aagtggcgac aacaacaaca acaacaacac caacaccatc
     6901 aacctcatcg taaccacagg cggcaccggc ttcgccgtgt ccgaccaaac caccgaagcc
     6961 gtctcggctc tcctacacaa gaatgctcct ggcctggtcc acggcatgct ggcggcttcg
     7021 ctgcaagtga ccccctttgc catgatgtca cgacctgtgg ctggcgtcag gaacggcacc
     7081 gttatcgttg cgctgcctgg gtcccccaag ggcgcgaagg agaatctgca agcagtgatc
     7141 aaaatgctgc cccacgcttg catgcaggct gctggtgcta actctagata agaattcgat
     7201 atcaagctta tcgataccgt cgacctcgag ggggggcccg gtaccagctt ttgttccctt
     7261 tagtgagggt taatagggcg tgcacggcta tggggtctga atgctaaaag acaccatttc
     7321 ccacactccc tctgattttt tttatggttt ttttttatct tcatttttcg ccgtctttta
     7381 ttcggttgtt tgggaggttc acatgcaacg tgcaagactg caagtatatt gagatgggcg
     7441 cttcatgcaa tgcagcatca ctgcagcgac attgtgacat ccacgtttcg atcgtctgtg
     7501 cttccccgtt catgtacgca tgatctgaaa aaaaaaaaaa aaaaaaaaag ttgatgtaat
     7561 aatgaaaata aacacccagg gacgagcacc ccttctggaa tagggagaaa aactaaaggg
     7621 acgtttgggt ctctacactt accaacagat tcatgtactg atcttcagcc tgtacagaac
     7681 agtacaatgt ccaaaactcg ttatcattac tctatgtgcc caaaatgacc cagatataca
     7741 tcaacccact acatcctttt gttcttgttt tttttttttt ttttgctact tcgaaaagaa
     7801 tttccgttta gttgctcttg ctcatgtgct caagacggat gctgaccgcc cttctgtgag
     7861 cctcgagctc ctcaacctta gccaactgca tgacagcctg gccgacgttt ttgagaccct
     7921 cggcagtcaa gttggagctg gtaatgtgct tgacgaacga ggcgagattg acgccagagt
     7981 actgcttggc aaagccatag gtagctatgt gtcttgacgt tagcagaaga atctgtgaag
     8041 cggggctatt tatgatatgt aacttacgca gcgagtggtt aacaccagca gagtaatcgc
     8101 caacggactc aggagtccag gcgccaatga agacactacc ggcgttcatg acaagatcga
     8161 cagccttctc ggcctccttg atctggagga tcaagtgctc aggagcgtac ttgttgctga
     8221 gctccatggc ctcctcgacg gtcttgacct gcacggtgat cgagtgggcg atggagccac
     8281 ggacaatctg gacgcgagga agctccgtag cctgacggtg aacctcgtcc tcaatagcct
     8341 gaagatgctc ctcgtcgagg tcaatagcga tcaggatgac ctgactgtca acgccgtgct
     8401 cagcctggga caggagatcc gaggcaacga acgcggggtt ggcgtccttg tcagcgatga
     8461 ccagcacctc ggacgggcca gcgggcatgt caataccaac ggcagcgttg gtgtcgttgc
     8521 tgacgaacat cttggcagca gtgacgaact ggttaccggg gccgagaatc ttgtcgacct
     8581 tggtgatgct ctcggtgccg taggccatgg cagctacggc ctgggcaccg ccggcaagca
     8641 cgatggactc ggcgccaacc ttgtgagcga cgtagacaat ctcgggagtg atggttccgt
     8701 cggcgcgggg aggagaggcg aacacaatct tgttgcagcc ggcgaccatg gcgggaacac
     8761 ccagcataag ggcagtgctg gggagaacgg cggtaccgcc ggggatgtag cagccgacgg
     8821 cctcgatggg acgagagaaa cggctgcaga caacaccggg catggtctcg acctggaggg
     8881 gcttctcctc cttctgggcg gcgtggaact tgcggatgtt ctcgaaggac acgtcgatgg
     8941 cagcaatggt ctcctcaggg agctgcataa gctccttggg gaagggcgcc ttcaggacgg
     9001 ggctagtaag agaggtagcc ttctcgaact tgtgagtgta cgacagaaca gccttgtcgc
     9061 cgttcttgcg gacgtcctcg atgatgggga caatgatctt gtagatggca tcggacgact
     9121 tttgcgcagg acgcttgaga gcagcatcga gctcctcggt agagaccttg gaggcgtcga
     9181 aacgtctcat ggtgatcttc tcaggggtgg tctcctgggc ggcctccttg gtgacagggg
     9241 ccgaagtggc agcgagagcg gacgccgcag gcttgatgcc ctccttctca gcccacttac
     9301 ccttagcatc tccagtcctg cgcttgacct tccagctctt ggcgtcaagg ctcctttcga
     9361 tatcggcaag agtaacgccg gcagcaacgg ccctggtaag agcaaagtag aagagatcgg
     9421 cagcctcaaa ggcgatttcc tggggggtct gagcggtgca gagctcctca gcctcctcca
     9481 tgatcttggc ccggactagc ttctcatcgg agaagagacg ggcagtgtag gagccctcgg
     9541 gggcagactg tttcctcgaa atcaaagtct gctcgagctt gggaaggcct ttgagctgac
     9601 caaagcagcc ggactgatcg aggtggcaga aacgaccctt ctgcttcacg acaaacttga
     9661 gagcatcgtt atcgcagtca agcgagatgc ggacgagctc ctgagtgtct ccggaagtag
     9721 cacccttgta ccagagaccg cgcttccggc tctgatagac accagtctgt gtcctgaggg
     9781 cctcgttcac actctcggca ctgctgtaga ccagacccag agcagtgtcg tgctcatcaa
     9841 cgacaacggt ggggagaaga ccatcgggac ggtcagactt ccagacgctc gagaggatgg
     9901 tggagatggc aagcttcagc tgctcgagtt c
//
