LOCUS       sequence-459452-        9385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NIT-9E with linkage region at the N terminus in N. 
            crassa at the his-3 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9385)
  TITLE     Convergent Evolution Links Molybdenum Insertase Domains with 
            Organism-Specific Sequences.
REFERENCE   2  (bases 1 to 9385)
  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_linker_nit9E
FEATURES             Location/Qualifiers
     source          1..9385
                     /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(6666..6682)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6667..6683)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(join(9384..9385,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(9385,1..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 atggctttac ataagggggg agtgaagaaa
     5041 ttggagcagg aggcgggagt agaaggaggt ggtggacgtg gacatggtca cggacatgga
     5101 cacggacatg gacacaagca tcacgaccat ggcgagcacc gaggaggagg agggggaggc
     5161 tgtggcgccc atgggcatgg ccataaccac ggtcacggcc caggccacgg ccacgccggt
     5221 ctagtccgac acaccatccc ctctgaaaac cccaagtcca acgatcccag cttaggtccc
     5281 agccgccgcc accgcgagtc tccttacccg atgctctctg tctccgaagc gctctcccaa
     5341 atagcgcaac acacgccgat gccgcaaatc gtcacctcca aagtcgatag ctcgctcatc
     5401 ggcgccgttc tggcgcagga cgtttcggcg cgcgaaaatg tgcccgcttt tcgcgctagc
     5461 atcgtcgacg gctacgccgt agtcgtgccc caagacggga acatgcgcgg cgtgttcccc
     5521 gtcgtgtcgg tttcgcatgc cgctcctgga gatggggaaa aaatgccgac gctcaaagag
     5581 ggcgagattg cccggatcac cacgggtgca cccctgcctc cgggtgcgac atcggttatt
     5641 atggtggagg atacggtgtt ggtgtcgacg acgtcatcca agacggagga tgcggaggat
     5701 ggaaagggag tggaggagga gaaggaggtg gagattttag cagctgataa agttcgcgaa
     5761 ggcgaaaatg ttagggaggt gggaagtgat atcaagcagg ggcaactgat tctaggcaag
     5821 ggagatcagg tgtctgctgt ggggggtgag attggtctgt tggccgccgt gggtgtgggt
     5881 gaggtgcagg tgtaccgacg tcctgttgtt ggcgttctat ctacgggtga tgagattgtg
     5941 gaccatgatc gcccgggaga cttgcggctg ggcgaggtgc gcgatacgaa ccggccaact
     6001 ctcatttcgg cggcgagaga ttgggggtat gaggtgattg atcttggagt ggcgcgggac
     6061 aaggccggga ctttggagga gacgctgcgg caggcattga ggcaggttga cgtggtgatc
     6121 acgaccggcg gagtgtccat gggtgagctg gatctcctca agccgacgat agagcgttcg
     6181 ctgggcggaa caattcgctt tggtcgggtg gcaatgaagc ccggcaagcc taccacgttt
     6241 gctaccgttc cggtaaagga taacgacgga gagagagttt ccaaggtaat cttctcgcta
     6301 ccaggtaacc cggcatcggc tttggttacg tttcacttgt ttgtactgcc gtctctgcac
     6361 caaatgtcgg gcgtctcacc acctggcttg ccaaaggtgc ctgtgacact gggccatgat
     6421 ttctccctcg atagatctag acccgagtac caccgggcca ttgtctccgt tagcagggac
     6481 ggagtgctca ccgcggtcag cacaggagga cagagaagct cgcgggtagg aagtatgaag
     6541 ggggccaacg cgcttctctg catgccttgc ggaccagagc cgttgcgcaa gggtgccaag
     6601 gtggaagcgt tgcttatggg cggtttacgc agcgatgtga tttaagaatt cgatatcaag
     6661 cttatcgata ccgtcgacct cgaggggggg cccggtacca gcttttgttc cctttagtga
     6721 gggttaatag ggcgtgcacg gctatggggt ctgaatgcta aaagacacca tttcccacac
     6781 tccctctgat tttttttatg gttttttttt atcttcattt ttcgccgtct tttattcggt
     6841 tgtttgggag gttcacatgc aacgtgcaag actgcaagta tattgagatg ggcgcttcat
     6901 gcaatgcagc atcactgcag cgacattgtg acatccacgt ttcgatcgtc tgtgcttccc
     6961 cgttcatgta cgcatgatct gaaaaaaaaa aaaaaaaaaa aagttgatgt aataatgaaa
     7021 ataaacaccc agggacgagc accccttctg gaatagggag aaaaactaaa gggacgtttg
     7081 ggtctctaca cttaccaaca gattcatgta ctgatcttca gcctgtacag aacagtacaa
     7141 tgtccaaaac tcgttatcat tactctatgt gcccaaaatg acccagatat acatcaaccc
     7201 actacatcct tttgttcttg tttttttttt ttttttttgc tacttcgaaa agaatttccg
     7261 tttagttgct cttgctcatg tgctcaagac ggatgctgac cgcccttctg tgagcctcga
     7321 gctcctcaac cttagccaac tgcatgacag cctggccgac gtttttgaga ccctcggcag
     7381 tcaagttgga gctggtaatg tgcttgacga acgaggcgag attgacgcca gagtactgct
     7441 tggcaaagcc ataggtagct atgtgtcttg acgttagcag aagaatctgt gaagcggggc
     7501 tatttatgat atgtaactta cgcagcgagt ggttaacacc agcagagtaa tcgccaacgg
     7561 actcaggagt ccaggcgcca atgaagacac taccggcgtt catgacaaga tcgacagcct
     7621 tctcggcctc cttgatctgg aggatcaagt gctcaggagc gtacttgttg ctgagctcca
     7681 tggcctcctc gacggtcttg acctgcacgg tgatcgagtg ggcgatggag ccacggacaa
     7741 tctggacgcg aggaagctcc gtagcctgac ggtgaacctc gtcctcaata gcctgaagat
     7801 gctcctcgtc gaggtcaata gcgatcagga tgacctgact gtcaacgccg tgctcagcct
     7861 gggacaggag atccgaggca acgaacgcgg ggttggcgtc cttgtcagcg atgaccagca
     7921 cctcggacgg gccagcgggc atgtcaatac caacggcagc gttggtgtcg ttgctgacga
     7981 acatcttggc agcagtgacg aactggttac cggggccgag aatcttgtcg accttggtga
     8041 tgctctcggt gccgtaggcc atggcagcta cggcctgggc accgccggca agcacgatgg
     8101 actcggcgcc aaccttgtga gcgacgtaga caatctcggg agtgatggtt ccgtcggcgc
     8161 ggggaggaga ggcgaacaca atcttgttgc agccggcgac catggcggga acacccagca
     8221 taagggcagt gctggggaga acggcggtac cgccggggat gtagcagccg acggcctcga
     8281 tgggacgaga gaaacggctg cagacaacac cgggcatggt ctcgacctgg aggggcttct
     8341 cctccttctg ggcggcgtgg aacttgcgga tgttctcgaa ggacacgtcg atggcagcaa
     8401 tggtctcctc agggagctgc ataagctcct tggggaaggg cgccttcagg acggggctag
     8461 taagagaggt agccttctcg aacttgtgag tgtacgacag aacagccttg tcgccgttct
     8521 tgcggacgtc ctcgatgatg gggacaatga tcttgtagat ggcatcggac gacttttgcg
     8581 caggacgctt gagagcagca tcgagctcct cggtagagac cttggaggcg tcgaaacgtc
     8641 tcatggtgat cttctcaggg gtggtctcct gggcggcctc cttggtgaca ggggccgaag
     8701 tggcagcgag agcggacgcc gcaggcttga tgccctcctt ctcagcccac ttacccttag
     8761 catctccagt cctgcgcttg accttccagc tcttggcgtc aaggctcctt tcgatatcgg
     8821 caagagtaac gccggcagca acggccctgg taagagcaaa gtagaagaga tcggcagcct
     8881 caaaggcgat ttcctggggg gtctgagcgg tgcagagctc ctcagcctcc tccatgatct
     8941 tggcccggac tagcttctca tcggagaaga gacgggcagt gtaggagccc tcgggggcag
     9001 actgtttcct cgaaatcaaa gtctgctcga gcttgggaag gcctttgagc tgaccaaagc
     9061 agccggactg atcgaggtgg cagaaacgac ccttctgctt cacgacaaac ttgagagcat
     9121 cgttatcgca gtcaagcgag atgcggacga gctcctgagt gtctccggaa gtagcaccct
     9181 tgtaccagag accgcgcttc cggctctgat agacaccagt ctgtgtcctg agggcctcgt
     9241 tcacactctc ggcactgctg tagaccagac ccagagcagt gtcgtgctca tcaacgacaa
     9301 cggtggggag aagaccatcg ggacggtcag acttccagac gctcgagagg atggtggaga
     9361 tggcaagctt cagctgctcg agttc
//
