LOCUS       sequence-408184-        4774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSU20 with addition of CEN6-ARSH4-HIS3 cassette for S. cerevisiae 
            and RK2 origin of conjugal transfer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4774)
  TITLE     Medium-copy shuttle vectors for E. coli, H. influenzae, and S. 
            cerevisiae with visual and antibiotic markers
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4774)
  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            "Unpublished"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: pHflu1
FEATURES             Location/Qualifiers
     source          1..4774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      37..582
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     483..502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     699..716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    953..974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(989..994,995..1012,1013..1019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1027..1043
                     /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     1032..1054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1051..1067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1051..1067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1063..1311
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAPRGVDSRGSPIDGVPSSNSLAVVLQRRDWENPGVT
                     QLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     primer_bind     complement(1149..1166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1149..1165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1158..1180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1555..1743
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1744..2406
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     misc_feature    2567..3075
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     oriT            3218..3327
                     /note="incP origin of transfer"
     CDS             3360..3731
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIE*KAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     promoter        3988..4090
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4091..4750
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4157..4176)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt
       61 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
      121 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
      181 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
      241 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
      361 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
      421 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
      481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
      541 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
      601 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
      661 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
      721 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
      781 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatatacac
      841 tccgctagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      901 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      961 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1021 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1081 agcttgcatg cccctcgagg ggtcgactct agaggatccc ccatcgatgg ggtaccgagc
     1141 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1201 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1261 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctg atgtccggcg
     1321 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa
     1381 acagaagcca ctggagcacc tcaaaaacac catcatacac taaatcatta attaataaga
     1441 gcttggtgag cgctaggagt cactgccagg tatcgtttga acacggcatt agtcagggaa
     1501 gtcataacac agtcctttcc cgcaattttc tttttctatt actcttggcc tcctctagta
     1561 cactctatat ttttttatgc ctcggtaatg attttcattt tttttttttc cacctagcgg
     1621 atgactcttt ttttttctta gcgattggca ttatcacata atgaattata cattatataa
     1681 agtaatgtga tttcttcgaa gaatatacta aaaaatgagc aggcaagata aacgaaggca
     1741 aagatgacag agcagaaagc cctagtaaag cgtattacaa atgaaaccaa gattcagatt
     1801 gcgatctctt taaagggtgg tcccctagcg atagagcact cgatcttccc agaaaaagag
     1861 gcagaagcag tagcagaaca ggccacacaa tcgcaagtga ttaacgtcca cacaggtata
     1921 gggtttctgg accatatgat acatgctctg gccaagcatt ccggctggtc gctaatcgtt
     1981 gagtgcattg gtgacttaca catagacgac catcacacca ctgaagactg cgggattgct
     2041 ctcggtcaag cttttaaaga ggccctaggg gccgtgcgtg gagtaaaaag gtttggatca
     2101 ggatttgcgc ctttggatga ggcactttcc agagcggtgg tagatctttc gaacaggccg
     2161 tacgcagttg tcgaacttgg tttgcaaagg gagaaagtag gagatctctc ttgcgagatg
     2221 atcccgcatt ttcttgaaag ctttgcagag gctagcagaa ttaccctcca cgttgattgt
     2281 ctgcgaggca agaatgatca tcaccgtagt gagagtgcgt tcaaggctct tgcggttgcc
     2341 ataagagaag ccacctcgcc caatggtacc aacgatgttc cctccaccaa aggtgttctt
     2401 atgtagtgac accgattatt taaagctgca gcatacgata tatatacatg tgtatatatg
     2461 tatacctatg aatgtcagta agtatgtata cgaacagtat gatactgaag atgacaaggt
     2521 aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc tttcctatca cgtgctataa
     2581 aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa
     2641 aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaatctaga gactctaggg
     2701 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     2761 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     2821 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     2881 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     2941 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     3001 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     3061 tacaggcaag cgatccacta gtatcttccg ctgcataacc ctgcttcggg gtcattatag
     3121 cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc
     3181 gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc
     3241 cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg
     3301 ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga
     3361 tggctgatga aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc
     3421 cagacgaacg aagagcgatt gagtaaaagg cggcggcggc cggcatgagc ctgtcggcct
     3481 acctgctggc cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc
     3541 gcgagctggc ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc
     3601 tcaccgacga cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga
     3661 agatcgaaga gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg
     3721 cagagccatg acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac
     3781 gtccccatgc gctccatcaa gaagaggcac ttcgagctgt aagtacatca ccgacgagca
     3841 aggcaagacg atcgagctag catcacccga cgcactttgc gccgaataaa tacctgtgac
     3901 ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg
     3961 ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca actttcacca
     4021 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     4081 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     4141 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     4201 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     4261 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     4321 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     4381 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     4441 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     4501 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     4561 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     4621 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     4681 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     4741 cggggcgtaa tttttttaag gcagttattg gtgc
//
