LOCUS       sequence-239185-        6002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-NatMx ICE plasmid used to facilitate yeast recombination 
            cloning and stable genomic Integration after CEN Excision..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6002)
  AUTHORS   Flagg MP, Kao A, Hampton RY
  TITLE     ICE plasmids: combining the ease of yeast recombination cloning with
            the stability of genomic integration.
  JOURNAL   Yeast. 2019 May 10. doi: 10.1002/yea.3400.
  PUBMED    31074531
REFERENCE   2  (bases 1 to 6002)
  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: "Yeast. 2019
            May 10. doi: 10.1002/yea.3400."
COMMENT     Sequence Label: pICE TRP1-NatMx
FEATURES             Location/Qualifiers
     source          1..6002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..101
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        122..403
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             404..1078
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1175..1630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1312..1333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1524..1543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1756..1778
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1770..1787
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1771..1787
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1797..1816
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1797..1815
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1840..1856
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1841..1857
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1891..1907)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1891..1907)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    1938..2441
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2496..2516)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2496..2514)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2535..2551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2535..2551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2548..2570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2559..2575
                     /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)."
     promoter        complement(join(2583..2589,2590..2607,2608..2613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2628..2649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2766..2783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2937..3525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3017..3036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3696..4487,4488..4556))
                     /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     4319..4338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4557..4661)
                     /gene="bla"
                     /label=AmpR promoter
     gene            4734..5853
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        4734..5077
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5078..5650
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      5656..5853
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     5906..5924
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5962..5984)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
       61 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatagat
      121 caacgacatt actatatata taatatagga agcatttaat agaacagcat cgtaatatat
      181 gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt attgaaaaat
      241 agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc gattaagaat
      301 taattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt gactattgag
      361 cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgtctg ttattaattt
      421 cacaggtagt tctggtccat tggtgaaagt ttgcggcttg cagagcacag aggccgcaga
      481 atgtgctcta gattccgatg ctgacttgct gggtattata tgtgtgccca atagaaagag
      541 aacaattgac ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag
      601 ttcaggcact ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt
      661 ggctctggtc aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggca
      721 agaataccaa gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg
      781 caacatacta ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc
      841 agaagcaggt gggacaggtg aacttttgga ttggaactcg atttctgact gggttggaag
      901 gcaagagagc cccgaaagct tacattttat gttagctggt ggactgacgc cagaaaatgt
      961 tggtgatgcg cttagattaa atggcgttat tggtgttgat gtaagcggag gtgtggagac
     1021 aaatggtgta aaagactcta acaaaatagc aaatttcgtc aaaaatgcta agaaataggt
     1081 tattactgag tagtatttat ttaattatat cagttattac cctatgcggt gtgaaatacc
     1141 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     1201 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1261 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     1321 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1381 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     1441 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     1501 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     1561 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     1621 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1681 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1741 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1801 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata
     1861 agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg gccgccctct
     1921 tagcgcttct taggacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg
     1981 atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt
     2041 tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg
     2101 tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag
     2161 attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg
     2221 tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga
     2281 gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa
     2341 caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt atatatttat
     2401 attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggcg
     2461 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2521 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2581 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2641 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2701 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2761 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2821 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2881 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2941 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3001 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3061 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3361 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4561 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4621 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4681 ccacctgacg tcttgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg
     4741 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     4801 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     4861 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     4921 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     4981 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     5041 gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaccactc ttgacgacac
     5101 ggcttaccgg taccgcacca gtgtcccggg ggacgccgag gccatcgagg cactggatgg
     5161 gtccttcacc accgacaccg tcttccgcgt caccgccacc ggggacggct tcaccctgcg
     5221 ggaggtgccg gtggacccgc ccctgaccaa ggtgttcccc gacgacgaat cggacgacga
     5281 atcggacgac ggggaggacg gcgacccgga ctcccggacg ttcgtcgcgt acggggacga
     5341 cggcgacctg gcgggcttcg tggtcgtctc gtactccggc tggaaccgcc ggctgaccgt
     5401 cgaggacatc gaggtcgccc cggagcaccg ggggcacggg gtcgggcgcg cgttgatggg
     5461 gctcgcgacg gagttcgccc gcgagcgggg cgccgggcac ctctggctgg aggtcaccaa
     5521 cgtcaacgca ccggcgatcc acgcgtaccg gcggatgggg ttcaccctct gcggcctgga
     5581 caccgccctg tacgacggca ccgcctcgga cggcgagcag gcgctctaca tgagcatgcc
     5641 ctgcccctaa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     5701 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     5761 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     5821 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     5881 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5941 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6001 tc
//
