LOCUS       sequence-6933-ex        6098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  TITLE     Cross lab plasmids
REFERENCE   2  (bases 1 to 6098)
  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"
COMMENT     Sequence Label: pRS420
FEATURES             Location/Qualifiers
     source          1..6098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            complement(249..1605)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(249..446)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(452..1261)
                     /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     1175..1194
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1262..1605)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     rep_origin      complement(1722..2177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1859..1880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1960..2538)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     2071..2090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2307..2329
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2321..2338
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2322..2338
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2348..2367
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2348..2366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2375..2482
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2391..2407
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2392..2408
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2442..2458)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2442..2458)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2495..2515)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2495..2513)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2534..2550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2534..2550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2547..2569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2558..2574
                     /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(2582..2588,2589..2606,2607..2612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2627..2648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2765..2782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2936..3524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3016..3035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3695..4486,4487..4555))
                     /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     4318..4337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4556..4660)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4687..6029
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5650..5697)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6071..6089
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataatca tcgatgaatt cgagctcgtt ttcgacactg gatggcggcg ttagtatcga
      241 atcgacagca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc
      301 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
      361 acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
      421 aaaacaagaa tctttttatt gtcagtactg attagaaaaa ctcatcgagc atcaaatgaa
      481 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
      541 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
      601 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
      661 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
      721 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
      781 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
      841 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
      901 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttgc
      961 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1021 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1081 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1141 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1201 aatcagcatc catgttggaa tttaatcgcg gcctcgaaac gtgagtcttt tccttaccca
     1261 tggttgttta tgttcggatg tgatgtgaga actgtatcct agcaagattt taaaaggaag
     1321 tatatgaaag aagaacctca gtggcaaatc ctaacctttt atatttctct acaggggcgc
     1381 ggcgtgggga caattcaacg cgtctgtgag gggagcgttt ccctgctcgc aggtctgcag
     1441 cgaggagccg taatttttgc ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga
     1501 ttatacatgg ggatgtatgg gctaaatgta cgggcgacag tcacatcatg cccctgagct
     1561 gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc
     1621 cggcggggac gaggcaagct aaacagatct ggcgcgcctt aattaaccct atgcggtgtg
     1681 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa acgttaatat
     1741 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1801 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1861 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1921 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1981 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2041 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2101 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2161 gccgctacag ggcgcgtcgc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     2221 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     2281 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     2341 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcga
     2401 cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc
     2461 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     2521 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2581 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2641 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2701 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2761 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2821 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2881 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2941 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3001 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3061 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3121 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3181 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3241 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3301 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3361 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3421 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3481 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3541 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3601 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3661 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3721 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3781 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3841 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3901 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3961 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4021 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4081 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4141 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4201 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4261 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4321 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4381 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4441 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4501 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4561 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4621 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4681 cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     4741 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     4801 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     4861 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     4921 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     4981 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     5041 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     5101 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     5161 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     5221 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     5281 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     5341 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     5401 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     5461 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     5521 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     5581 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa
     5641 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa
     5701 gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact
     5761 gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc
     5821 atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg
     5881 tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca
     5941 gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc
     6001 aatgctatca tttcctttga tattggatca tatctaagaa accattatta tcatgacatt
     6061 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
