LOCUS       sequence-459573-        9881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NIT-9 with moco resulfurase 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 9881)
  TITLE     Convergent Evolution Links Molybdenum Insertase Domains with 
            Organism-Specific Sequences.
REFERENCE   2  (bases 1 to 9881)
  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_random_linker
FEATURES             Location/Qualifiers
     source          1..9881
                     /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(7158..7174)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7159..7175)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(join(9880..9881,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(9881,1..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 aactctagag actggcctgt ccctcaggcc
     5581 catgctgaca ccggggagac tggagccaca tcagcagaca gccaggctga tgttatacct
     5641 gctgtcatgg gcagacgtag cctctcgcct caggaagatg ccctcacagg ctccagggtt
     5701 tggaacaact cgtctactgt gaatgctgtg cctgtggccc cacctgtgtg tgatgtcgcc
     5761 agaacccagc cgactccttc agagaaagct gcaggagtcc tgatgctctc tgtctccgaa
     5821 gcgctctccc aaatagcgca acacacgccg atgccgcaaa tcgtcacctc caaagtcgat
     5881 agctcgctca tcggcgccgt tctggcgcag gacgtttcgg cgcgcgaaaa tgtgcccgct
     5941 tttcgcgcta gcatcgtcga cggctacgcc gtagtcgtgc cccaagacgg gaacatgcgc
     6001 ggcgtgttcc ccgtcgtgtc ggtttcgcat gccgctcctg gagatgggga aaaaatgccg
     6061 acgctcaaag agggcgagat tgcccggatc accacgggtg cacccctgcc tccgggtgcg
     6121 acatcggtta ttatggtgga ggatacggtg ttggtgtcga cgacgtcatc caagacggag
     6181 gatgcggagg atggaaaggg agtggaggag gagaaggagg tggagatttt agcagctgat
     6241 aaagttcgcg aaggcgaaaa tgttagggag gtgggaagtg atatcaagca ggggcaactg
     6301 attctaggca agggagatca ggtgtctgct gtggggggtg agattggtct gttggccgcc
     6361 gtgggtgtgg gtgaggtgca ggtgtaccga cgtcctgttg ttggcgttct atctacgggt
     6421 gatgagattg tggaccatga tcgcccggga gacttgcggc tgggcgaggt gcgcgatacg
     6481 aaccggccaa ctctcatttc ggcggcgaga gattgggggt atgaggtgat tgatcttgga
     6541 gtggcgcggg acaaggccgg gactttggag gagacgctgc ggcaggcatt gaggcaggtt
     6601 gacgtggtga tcacgaccgg cggagtgtcc atgggtgagc tggatctcct caagccgacg
     6661 atagagcgtt cgctgggcgg aacaattcgc tttggtcggg tggcaatgaa gcccggcaag
     6721 cctaccacgt ttgctaccgt tccggtaaag gataacgacg gagagagagt ttccaaggta
     6781 atcttctcgc taccaggtaa cccggcatcg gctttggtta cgtttcactt gtttgtactg
     6841 ccgtctctgc accaaatgtc gggcgtctca ccacctggct tgccaaaggt gcctgtgaca
     6901 ctgggccatg atttctccct cgatagatct agacccgagt accaccgggc cattgtctcc
     6961 gttagcaggg acggagtgct caccgcggtc agcacaggag gacagagaag ctcgcgggta
     7021 ggaagtatga agggggccaa cgcgcttctc tgcatgcctt gcggaccaga gccgttgcgc
     7081 aagggtgcca aggtggaagc gttgcttatg ggcggtttac gcagcgatgt gatttaagaa
     7141 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac cagcttttgt
     7201 tccctttagt gagggttaat agggcgtgca cggctatggg gtctgaatgc taaaagacac
     7261 catttcccac actccctctg atttttttta tggttttttt ttatcttcat ttttcgccgt
     7321 cttttattcg gttgtttggg aggttcacat gcaacgtgca agactgcaag tatattgaga
     7381 tgggcgcttc atgcaatgca gcatcactgc agcgacattg tgacatccac gtttcgatcg
     7441 tctgtgcttc cccgttcatg tacgcatgat ctgaaaaaaa aaaaaaaaaa aaaaaagttg
     7501 atgtaataat gaaaataaac acccagggac gagcacccct tctggaatag ggagaaaaac
     7561 taaagggacg tttgggtctc tacacttacc aacagattca tgtactgatc ttcagcctgt
     7621 acagaacagt acaatgtcca aaactcgtta tcattactct atgtgcccaa aatgacccag
     7681 atatacatca acccactaca tccttttgtt cttgtttttt tttttttttt ttttgctact
     7741 tcgaaaagaa tttccgttta gttgctcttg ctcatgtgct caagacggat gctgaccgcc
     7801 cttctgtgag cctcgagctc ctcaacctta gccaactgca tgacagcctg gccgacgttt
     7861 ttgagaccct cggcagtcaa gttggagctg gtaatgtgct tgacgaacga ggcgagattg
     7921 acgccagagt actgcttggc aaagccatag gtagctatgt gtcttgacgt tagcagaaga
     7981 atctgtgaag cggggctatt tatgatatgt aacttacgca gcgagtggtt aacaccagca
     8041 gagtaatcgc caacggactc aggagtccag gcgccaatga agacactacc ggcgttcatg
     8101 acaagatcga cagccttctc ggcctccttg atctggagga tcaagtgctc aggagcgtac
     8161 ttgttgctga gctccatggc ctcctcgacg gtcttgacct gcacggtgat cgagtgggcg
     8221 atggagccac ggacaatctg gacgcgagga agctccgtag cctgacggtg aacctcgtcc
     8281 tcaatagcct gaagatgctc ctcgtcgagg tcaatagcga tcaggatgac ctgactgtca
     8341 acgccgtgct cagcctggga caggagatcc gaggcaacga acgcggggtt ggcgtccttg
     8401 tcagcgatga ccagcacctc ggacgggcca gcgggcatgt caataccaac ggcagcgttg
     8461 gtgtcgttgc tgacgaacat cttggcagca gtgacgaact ggttaccggg gccgagaatc
     8521 ttgtcgacct tggtgatgct ctcggtgccg taggccatgg cagctacggc ctgggcaccg
     8581 ccggcaagca cgatggactc ggcgccaacc ttgtgagcga cgtagacaat ctcgggagtg
     8641 atggttccgt cggcgcgggg aggagaggcg aacacaatct tgttgcagcc ggcgaccatg
     8701 gcgggaacac ccagcataag ggcagtgctg gggagaacgg cggtaccgcc ggggatgtag
     8761 cagccgacgg cctcgatggg acgagagaaa cggctgcaga caacaccggg catggtctcg
     8821 acctggaggg gcttctcctc cttctgggcg gcgtggaact tgcggatgtt ctcgaaggac
     8881 acgtcgatgg cagcaatggt ctcctcaggg agctgcataa gctccttggg gaagggcgcc
     8941 ttcaggacgg ggctagtaag agaggtagcc ttctcgaact tgtgagtgta cgacagaaca
     9001 gccttgtcgc cgttcttgcg gacgtcctcg atgatgggga caatgatctt gtagatggca
     9061 tcggacgact tttgcgcagg acgcttgaga gcagcatcga gctcctcggt agagaccttg
     9121 gaggcgtcga aacgtctcat ggtgatcttc tcaggggtgg tctcctgggc ggcctccttg
     9181 gtgacagggg ccgaagtggc agcgagagcg gacgccgcag gcttgatgcc ctccttctca
     9241 gcccacttac ccttagcatc tccagtcctg cgcttgacct tccagctctt ggcgtcaagg
     9301 ctcctttcga tatcggcaag agtaacgccg gcagcaacgg ccctggtaag agcaaagtag
     9361 aagagatcgg cagcctcaaa ggcgatttcc tggggggtct gagcggtgca gagctcctca
     9421 gcctcctcca tgatcttggc ccggactagc ttctcatcgg agaagagacg ggcagtgtag
     9481 gagccctcgg gggcagactg tttcctcgaa atcaaagtct gctcgagctt gggaaggcct
     9541 ttgagctgac caaagcagcc ggactgatcg aggtggcaga aacgaccctt ctgcttcacg
     9601 acaaacttga gagcatcgtt atcgcagtca agcgagatgc ggacgagctc ctgagtgtct
     9661 ccggaagtag cacccttgta ccagagaccg cgcttccggc tctgatagac accagtctgt
     9721 gtcctgaggg cctcgttcac actctcggca ctgctgtaga ccagacccag agcagtgtcg
     9781 tgctcatcaa cgacaacggt ggggagaaga ccatcgggac ggtcagactt ccagacgctc
     9841 gagaggatgg tggagatggc aagcttcagc tgctcgagtt c
//
