LOCUS       sequence-90633-e        7031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN vector with clonNat resistance marker for single Gateway 
            insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7031)
  TITLE     Fluorescent X-QTL
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7031)
  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            "Unpublished"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 1-F GW
FEATURES             Location/Qualifiers
     source          1..7031
                     /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    2853..2977
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3002..3007,3008..3025,3026..3032)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3086..3745
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3152..3171)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4090..4395
                     /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     4274..4293
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4436..4560)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4609..4625)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4610..4626)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4628..4875
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4644..4662)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4893..4912)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4894..4912)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4922..4939)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4922..4938)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4931..4953)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5083..5538
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5170..5189)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5380..5401
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            complement(5668..6787)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     terminator      complement(5668..5865)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(5871..6443)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(6444..6787)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(6862..6881)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6981..7003
                     /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 ccgaactagt ggatccccca tcacaagttt gtacaaaaaa gctgaacgag
     2881 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     2941 cataatactg taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg
     3001 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag
     3061 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     3121 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     3181 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     3241 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     3301 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     3361 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     3421 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     3481 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     3541 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     3601 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     3661 tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     3721 cgatgagtgg cagggcgggg cgtaaacgcc gcgtggatcc ggcttactaa aagccagata
     3781 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     3841 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     3901 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     3961 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     4021 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     4081 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     4141 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     4201 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     4261 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     4321 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     4381 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag
     4441 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     4501 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt
     4561 gatgggctgc aggaattcgg ggctgcagga attcgatatc aagcttatcg ataccgtcga
     4621 cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct
     4681 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
     4741 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     4801 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     4861 aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     4921 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4981 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5041 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc tgtagcggcg
     5101 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5161 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5221 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5281 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5341 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5401 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5461 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5521 tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5581 acaccgcata gctagtggat ctgatatcat cgatgaattc gagctcgttt tcgacactgg
     5641 atggcggcgt tagtatcgaa tcgacagcag tatagcgacc agcattcaca tacgattgac
     5701 gcatgatatt actttctgcg cacttaactt cgcatctggg cagatgatgt cgaggcgaaa
     5761 aaaaatataa atcacgctaa catttgatta aaatagaaca actacaatat aaaaaaacta
     5821 tacaaatgac aagttcttga aaacaagaat ctttttattg tcagtactga ttaggggcag
     5881 ggcatgctca tgtagagcgc ctgctcgccg tccgaggcgg tgccgtcgta cagggcggtg
     5941 tccaggccgc agagggtgaa ccccatccgc cggtacgcgt ggatcgccgg tgcgttgacg
     6001 ttggtgacct ccagccagag gtgcccggcg ccccgctcgc gggcgaactc cgtcgcgagc
     6061 cccatcaacg cgcgcccgac cccgtgcccc cggtgctccg gggcgacctc gatgtcctcg
     6121 acggtcagcc ggcggttcca gccggagtac gagacgacca cgaagcccgc caggtcgccg
     6181 tcgtccccgt acgcgacgaa cgtccgggag tccgggtcgc cgtcctcccc gtcgtccgat
     6241 tcgtcgtccg attcgtcgtc ggggaacacc ttggtcaggg gcgggtccac cggcacctcc
     6301 cgcagggtga agccgtcccc ggtggcggtg acgcggaaga cggtgtcggt ggtgaaggac
     6361 ccatccagtg cctcgatggc ctcggcgtcc cccgggacac tggtgcggta ccggtaagcc
     6421 gtgtcgtcaa gagtggtacc catggttgtt tatgttcgga tgtgatgtga gaactgtatc
     6481 ctagcaagat tttaaaagga agtatatgaa agaagaacct cagtggcaaa tcctaacctt
     6541 ttatatttct ctacaggggc gcggcgtggg gacaattcaa cgcgtctgtg aggggagcgt
     6601 ttccctgctc gcaggtctgc agcgaggagc cgtaattttt gcttcgcgcc gtgcggccat
     6661 caaaatgtat ggatgcaaat gattatacat ggggatgtat gggctaaatg tacgggcgac
     6721 agtcacatca tgcccctgag ctgcgcacgt caagactgtc aaggagggta ttctgggcct
     6781 ccatgtcgct ggccgggtga cccggcgggg acaaggcaag ctaaacagat cttgaaaagc
     6841 tgtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6901 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6961 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7021 gaaacgcgcg a
//
