LOCUS       sequence-90658-e        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN vector with G418 resistance marker for triple Gateway insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  TITLE     Fluorescent X-QTL
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7715)
  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            "Unpublished"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: p41Neo 3-F GW
FEATURES             Location/Qualifiers
     source          1..7715
                     /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      5072..5319
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5088..5106)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5337..5356)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5338..5356)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5366..5382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5375..5397)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5527..5982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5614..5633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5824..5845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            complement(6112..7470)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(6112..6309)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(6315..7124)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7038..7057
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(7127..7470)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(7546..7565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7665..7687
                     /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 ctcgagtcga gtcatgtaat tagttatgtc acgcttacat
     5101 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     5161 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     5221 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     5281 cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc aattcgccct
     5341 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5401 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5461 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5521 ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5581 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5641 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5701 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5761 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5821 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5881 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5941 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc ggtattttct
     6001 ccttacgcat ctgtgcggta tttcacaccg catagctagt ggatctgata tcatcgatga
     6061 attcgagctc gttttcgaca ctggatggcg gcgttagtat cgaatcgaca gcagtatagc
     6121 gaccagcatt cacatacgat tgacgcatga tattactttc tgcgcactta acttcgcatc
     6181 tgggcagatg atgtcgaggc gaaaaaaaat ataaatcacg ctaacatttg attaaaatag
     6241 aacaactaca atataaaaaa actatacaaa tgacaagttc ttgaaaacaa gaatcttttt
     6301 attgtcagta ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     6361 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     6421 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     6481 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     6541 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     6601 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     6661 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     6721 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     6781 cctgaatcag gatattcttc taatacctgg aatgctgttt tgccggggat cgcagtggtg
     6841 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     6901 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     6961 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     7021 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     7081 gaatttaatc gcggcctcga aacgtgagtc ttttccttac ccatatggtt gtttatgttc
     7141 ggatgtgatg tgagaactgt atcctagcaa gattttaaaa ggaagtatat gaaagaagaa
     7201 cctcagtggc aaatcctaac cttttatatt tctctacagg ggcgcggcgt ggggacaatt
     7261 caacgcgtct gtgaggggag cgtttccctg ctcgcaggtc tgcagcgagg agccgtaatt
     7321 tttgcttcgc gccgtgcggc catcaaaatg tatggatgca aatgattata catggggatg
     7381 tatgggctaa atgtacgggc gacagtcaca tcatgcccct gagctgcgca cgtcaagact
     7441 gtcaaggagg gtattctggg cctccatgtc gctggccggg tgacccggcg gggacaaggc
     7501 aagctaaaca agatcttgaa aagctgtggt atggtgcact ctcagtacaa tctgctctga
     7561 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     7621 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     7681 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
