LOCUS       sequence-90657-e        7470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN vector with clonNat resistance marker for triple Gateway 
            insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7470)
  TITLE     Fluorescent X-QTL
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7470)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: p41Nat 3-F GW
FEATURES             Location/Qualifiers
     source          1..7470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2812..2828
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2812..2828
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2849..2972
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3320..3625)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3422..3441)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(3945..4628)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     4543..4562
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4629..4731)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(4877..5000)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5048..5064)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5049..5065)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5067..5314
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5083..5101)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5332..5351)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5333..5351)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5361..5377)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5370..5392)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5522..5977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5609..5628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5819..5840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            complement(6107..7226)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     terminator      complement(6107..6304)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(6310..6882)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(6883..7226)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(7301..7320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7420..7442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaact
     2821 agtggatccc ccgattacgc caagctatca actttgtata gaaaagttga acgagaaacg
     2881 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     2941 tactgtaaaa cacaacatat ccagtcacta tggtcgaccg acagccttcc aaatgttctt
     3001 ctcaaacgga atcgtcgtat ccagcctact cgctattgtc ctcaatgccg tattaaatca
     3061 taaaaagaaa taagaaaaag aggtgcgagc ctcttttttg tgtgacaaaa taaaaacatc
     3121 tacctattca tatacgctag tgtcatagtc ctgaaaatca tctgcatcaa gaacaatttc
     3181 acaactctta tacttttctc ttacaagtcg ttcggcttca tctggatttt cagcctctat
     3241 acttactaaa cgtgataaag tttctgtaat ttctactgta tcgacctgca gactggctgt
     3301 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     3361 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
     3421 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     3481 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     3541 cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     3601 tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg
     3661 ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt
     3721 tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg
     3781 acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct
     3841 gcttcatagc atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat
     3901 accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc
     3961 acgcggcgtt tacgcccccc ctgccactca tcgcagtact gttgtaattc attaagcatt
     4021 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     4081 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     4141 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     4201 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     4261 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     4321 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     4381 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
     4441 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     4501 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     4561 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     4621 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     4681 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
     4741 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
     4801 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
     4861 gccaacttac ggccgccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg
     4921 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     4981 aactttatta tacatagttg ataattcact ggccgtcggg gctgcaggaa ttcgatatca
     5041 agcttatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg
     5101 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     5161 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     5221 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     5281 gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt
     5341 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5401 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5461 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5521 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5941 aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat tttctcctta
     6001 cgcatctgtg cggtatttca caccgcatag ctagtggatc tgatatcatc gatgaattcg
     6061 agctcgtttt cgacactgga tggcggcgtt agtatcgaat cgacagcagt atagcgacca
     6121 gcattcacat acgattgacg catgatatta ctttctgcgc acttaacttc gcatctgggc
     6181 agatgatgtc gaggcgaaaa aaaatataaa tcacgctaac atttgattaa aatagaacaa
     6241 ctacaatata aaaaaactat acaaatgaca agttcttgaa aacaagaatc tttttattgt
     6301 cagtactgat taggggcagg gcatgctcat gtagagcgcc tgctcgccgt ccgaggcggt
     6361 gccgtcgtac agggcggtgt ccaggccgca gagggtgaac cccatccgcc ggtacgcgtg
     6421 gatcgccggt gcgttgacgt tggtgacctc cagccagagg tgcccggcgc cccgctcgcg
     6481 ggcgaactcc gtcgcgagcc ccatcaacgc gcgcccgacc ccgtgccccc ggtgctccgg
     6541 ggcgacctcg atgtcctcga cggtcagccg gcggttccag ccggagtacg agacgaccac
     6601 gaagcccgcc aggtcgccgt cgtccccgta cgcgacgaac gtccgggagt ccgggtcgcc
     6661 gtcctccccg tcgtccgatt cgtcgtccga ttcgtcgtcg gggaacacct tggtcagggg
     6721 cgggtccacc ggcacctccc gcagggtgaa gccgtccccg gtggcggtga cgcggaagac
     6781 ggtgtcggtg gtgaaggacc catccagtgc ctcgatggcc tcggcgtccc ccgggacact
     6841 ggtgcggtac cggtaagccg tgtcgtcaag agtggtaccc atggttgttt atgttcggat
     6901 gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc
     6961 agtggcaaat cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac
     7021 gcgtctgtga ggggagcgtt tccctgctcg caggtctgca gcgaggagcc gtaatttttg
     7081 cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg
     7141 ggctaaatgt acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca
     7201 aggagggtat tctgggcctc catgtcgctg gccgggtgac ccggcgggga caaggcaagc
     7261 taaacagatc ttgaaaagct gtggtatggt gcactctcag tacaatctgc tctgatgccg
     7321 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     7381 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     7441 ggttttcacc gtcatcaccg aaacgcgcga
//
