LOCUS       sequence-69537-e        4343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Base vector for PCR-based DNA transplant in yeast. DNA segments to 
            be transplanted into the yeast genome are cloned into the MCS. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4343)
  AUTHORS   Horecka J, Chu AM, Davis RW
  TITLE     IpO: plasmids and methods for simplified, PCR-based DNA transplant 
            in yeast.
  JOURNAL   Yeast. 2014 May;31(5):185-93. doi: 10.1002/yea.3006. Epub 2014 Mar 
            20.
  PUBMED    24604451
REFERENCE   2  (bases 1 to 4343)
  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"; doi: "10.1002/yea.3006"; 
            journalName: "Yeast"; date: "2014-05"; volume: "31"; issue: "5"; 
            pages: "185-93"
COMMENT     Sequence Label: IpO
FEATURES             Location/Qualifiers
     source          1..4343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     679..695
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     679..695
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(729..745)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(730..746)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        773..989
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(1498..1517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1499..1517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1527..1544)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1527..1543)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1536..1558)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1684..2139)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1821..1842)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2033..2052
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2166..2270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2271..2339,2340..3131)
                     /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(2489..2508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3302..3890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3791..3810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4044..4061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4178..4199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4214..4219,4220..4237,4238..4244)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4252..4268
                     /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     4257..4279
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4276..4292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4276..4292
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4311..4331
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4313..4331
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa
       61 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
      121 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
      181 agcggtttga agcaggcggc ggaagaagta acaaaggaac ctagaggcct tttgatgtta
      241 gcagaattgt catgcaaggg ctccctagct actggagaat atactaaggg tactgttgac
      301 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
      361 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
      421 gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt
      481 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
      541 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
      601 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
      661 cagttattac ccggccgctc tagaactagt ggatcccccg ggctgcagga attcgatatc
      721 aagcttatcg ataccgtcga cctcgagtgt ggtttcaggg tccataaagc ttttcaattc
      781 atcttttttt tttttgttct tttttttgat tccggtttct ttgaaatttt tttgattcgg
      841 taatctccga gcagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
      901 gcatatgtgg tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
      961 caaaaacctg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1021 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1081 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1141 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     1201 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag
     1261 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     1321 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     1381 gcggtttgaa gcaggcggcg gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     1441 cagaattgtc atgcaagggc tccctagcta ctggagaata tactggtacc caattcgccc
     1501 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     1561 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     1621 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     1681 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1741 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1801 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1861 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1921 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1981 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2041 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2101 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2161 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2221 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2281 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2341 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2401 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2461 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2521 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2581 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2641 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2701 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2761 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2821 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2881 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2941 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3001 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3061 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3121 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3181 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3241 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3421 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3661 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3721 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3781 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3841 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3901 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3961 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4021 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4081 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4141 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4201 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4261 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     4321 ctcactaaag ggaacaaaag ctg
//
