LOCUS       sequence-459451-        9636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NIT-9 without linkage region in N. crassa at the his-3
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9636)
  TITLE     Convergent Evolution Links Molybdenum Insertase Domains with 
            Organism-Specific Sequences.
REFERENCE   2  (bases 1 to 9636)
  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_nit9_without_linker
FEATURES             Location/Qualifiers
     source          1..9636
                     /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(6915..6931)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6916..6932)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(join(9635..9636,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(9636,1..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 atgtcctcaa gtactactcc agcagcgctc
     5041 aaagcagcca tccttatcgt ttcaacgaca gcagctcaag accccgcaac ggactcggcc
     5101 gacgtcactc tccgccaggt cttcgatgaa caacaagccc aatgggaggt ccatgaatcc
     5161 aaaatcgttc ccgacgatgt cgtgcaaatc cagaagcaaa tcatccactg gacaaaccaa
     5221 gccaccgccg caagtggcga caacaacaac aacaacaaca ccaacaccat caacctcatc
     5281 gtaaccacag gcggcaccgg cttcgccgtg tccgaccaaa ccaccgaagc cgtctcggct
     5341 ctcctacaca agaatgctcc tggcctggtc cacggcatgc tggcggcttc gctgcaagtg
     5401 accccctttg ccatgatgtc acgacctgtg gctggcgtca ggaacggcac cgttatcgtt
     5461 gcgctgcctg ggtcccccaa gggcgcgaag gagaatctgc aagcagtgat caaaatgctg
     5521 ccccacgctt gcatgcaggc tgctggtgct aactctagaa tgctctctgt ctccgaagcg
     5581 ctctcccaaa tagcgcaaca cacgccgatg ccgcaaatcg tcacctccaa agtcgatagc
     5641 tcgctcatcg gcgccgttct ggcgcaggac gtttcggcgc gcgaaaatgt gcccgctttt
     5701 cgcgctagca tcgtcgacgg ctacgccgta gtcgtgcccc aagacgggaa catgcgcggc
     5761 gtgttccccg tcgtgtcggt ttcgcatgcc gctcctggag atggggaaaa aatgccgacg
     5821 ctcaaagagg gcgagattgc ccggatcacc acgggtgcac ccctgcctcc gggtgcgaca
     5881 tcggttatta tggtggagga tacggtgttg gtgtcgacga cgtcatccaa gacggaggat
     5941 gcggaggatg gaaagggagt ggaggaggag aaggaggtgg agattttagc agctgataaa
     6001 gttcgcgaag gcgaaaatgt tagggaggtg ggaagtgata tcaagcaggg gcaactgatt
     6061 ctaggcaagg gagatcaggt gtctgctgtg gggggtgaga ttggtctgtt ggccgccgtg
     6121 ggtgtgggtg aggtgcaggt gtaccgacgt cctgttgttg gcgttctatc tacgggtgat
     6181 gagattgtgg accatgatcg cccgggagac ttgcggctgg gcgaggtgcg cgatacgaac
     6241 cggccaactc tcatttcggc ggcgagagat tgggggtatg aggtgattga tcttggagtg
     6301 gcgcgggaca aggccgggac tttggaggag acgctgcggc aggcattgag gcaggttgac
     6361 gtggtgatca cgaccggcgg agtgtccatg ggtgagctgg atctcctcaa gccgacgata
     6421 gagcgttcgc tgggcggaac aattcgcttt ggtcgggtgg caatgaagcc cggcaagcct
     6481 accacgtttg ctaccgttcc ggtaaaggat aacgacggag agagagtttc caaggtaatc
     6541 ttctcgctac caggtaaccc ggcatcggct ttggttacgt ttcacttgtt tgtactgccg
     6601 tctctgcacc aaatgtcggg cgtctcacca cctggcttgc caaaggtgcc tgtgacactg
     6661 ggccatgatt tctccctcga tagatctaga cccgagtacc accgggccat tgtctccgtt
     6721 agcagggacg gagtgctcac cgcggtcagc acaggaggac agagaagctc gcgggtagga
     6781 agtatgaagg gggccaacgc gcttctctgc atgccttgcg gaccagagcc gttgcgcaag
     6841 ggtgccaagg tggaagcgtt gcttatgggc ggtttacgca gcgatgtgat ttaagaattc
     6901 gatatcaagc ttatcgatac cgtcgacctc gagggggggc ccggtaccag cttttgttcc
     6961 ctttagtgag ggttaatagg gcgtgcacgg ctatggggtc tgaatgctaa aagacaccat
     7021 ttcccacact ccctctgatt ttttttatgg ttttttttta tcttcatttt tcgccgtctt
     7081 ttattcggtt gtttgggagg ttcacatgca acgtgcaaga ctgcaagtat attgagatgg
     7141 gcgcttcatg caatgcagca tcactgcagc gacattgtga catccacgtt tcgatcgtct
     7201 gtgcttcccc gttcatgtac gcatgatctg aaaaaaaaaa aaaaaaaaaa aaagttgatg
     7261 taataatgaa aataaacacc cagggacgag caccccttct ggaataggga gaaaaactaa
     7321 agggacgttt gggtctctac acttaccaac agattcatgt actgatcttc agcctgtaca
     7381 gaacagtaca atgtccaaaa ctcgttatca ttactctatg tgcccaaaat gacccagata
     7441 tacatcaacc cactacatcc ttttgttctt gttttttttt tttttttttg ctacttcgaa
     7501 aagaatttcc gtttagttgc tcttgctcat gtgctcaaga cggatgctga ccgcccttct
     7561 gtgagcctcg agctcctcaa ccttagccaa ctgcatgaca gcctggccga cgtttttgag
     7621 accctcggca gtcaagttgg agctggtaat gtgcttgacg aacgaggcga gattgacgcc
     7681 agagtactgc ttggcaaagc cataggtagc tatgtgtctt gacgttagca gaagaatctg
     7741 tgaagcgggg ctatttatga tatgtaactt acgcagcgag tggttaacac cagcagagta
     7801 atcgccaacg gactcaggag tccaggcgcc aatgaagaca ctaccggcgt tcatgacaag
     7861 atcgacagcc ttctcggcct ccttgatctg gaggatcaag tgctcaggag cgtacttgtt
     7921 gctgagctcc atggcctcct cgacggtctt gacctgcacg gtgatcgagt gggcgatgga
     7981 gccacggaca atctggacgc gaggaagctc cgtagcctga cggtgaacct cgtcctcaat
     8041 agcctgaaga tgctcctcgt cgaggtcaat agcgatcagg atgacctgac tgtcaacgcc
     8101 gtgctcagcc tgggacagga gatccgaggc aacgaacgcg gggttggcgt ccttgtcagc
     8161 gatgaccagc acctcggacg ggccagcggg catgtcaata ccaacggcag cgttggtgtc
     8221 gttgctgacg aacatcttgg cagcagtgac gaactggtta ccggggccga gaatcttgtc
     8281 gaccttggtg atgctctcgg tgccgtaggc catggcagct acggcctggg caccgccggc
     8341 aagcacgatg gactcggcgc caaccttgtg agcgacgtag acaatctcgg gagtgatggt
     8401 tccgtcggcg cggggaggag aggcgaacac aatcttgttg cagccggcga ccatggcggg
     8461 aacacccagc ataagggcag tgctggggag aacggcggta ccgccgggga tgtagcagcc
     8521 gacggcctcg atgggacgag agaaacggct gcagacaaca ccgggcatgg tctcgacctg
     8581 gaggggcttc tcctccttct gggcggcgtg gaacttgcgg atgttctcga aggacacgtc
     8641 gatggcagca atggtctcct cagggagctg cataagctcc ttggggaagg gcgccttcag
     8701 gacggggcta gtaagagagg tagccttctc gaacttgtga gtgtacgaca gaacagcctt
     8761 gtcgccgttc ttgcggacgt cctcgatgat ggggacaatg atcttgtaga tggcatcgga
     8821 cgacttttgc gcaggacgct tgagagcagc atcgagctcc tcggtagaga ccttggaggc
     8881 gtcgaaacgt ctcatggtga tcttctcagg ggtggtctcc tgggcggcct ccttggtgac
     8941 aggggccgaa gtggcagcga gagcggacgc cgcaggcttg atgccctcct tctcagccca
     9001 cttaccctta gcatctccag tcctgcgctt gaccttccag ctcttggcgt caaggctcct
     9061 ttcgatatcg gcaagagtaa cgccggcagc aacggccctg gtaagagcaa agtagaagag
     9121 atcggcagcc tcaaaggcga tttcctgggg ggtctgagcg gtgcagagct cctcagcctc
     9181 ctccatgatc ttggcccgga ctagcttctc atcggagaag agacgggcag tgtaggagcc
     9241 ctcgggggca gactgtttcc tcgaaatcaa agtctgctcg agcttgggaa ggcctttgag
     9301 ctgaccaaag cagccggact gatcgaggtg gcagaaacga cccttctgct tcacgacaaa
     9361 cttgagagca tcgttatcgc agtcaagcga gatgcggacg agctcctgag tgtctccgga
     9421 agtagcaccc ttgtaccaga gaccgcgctt ccggctctga tagacaccag tctgtgtcct
     9481 gagggcctcg ttcacactct cggcactgct gtagaccaga cccagagcag tgtcgtgctc
     9541 atcaacgaca acggtgggga gaagaccatc gggacggtca gacttccaga cgctcgagag
     9601 gatggtggag atggcaagct tcagctgctc gagttc
//
