LOCUS       sequence-11497-e        4348 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 4348)
  AUTHORS   Schowalter RM, Pastrana DV, Pumphrey KA, Moyer AL, Buck CB.
  TITLE     Merkel cell polyomavirus and two previously unknown polyomaviruses 
            are chronically shed from human skin.
  JOURNAL   Cell Host Microbe. 2010 Jun 25;7(6):509-15.
  PUBMED    20542254
REFERENCE   2  (bases 1 to 4348)
  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: "Cell Host 
            Microbe."; date: "2010-06-25"; volume: "7(6)"; pages: "509-15"
COMMENT     Sequence Label: pFunnyfarm
FEATURES             Location/Qualifiers
     source          1..4348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     176..198
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     190..207
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     191..207
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    224..455
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(851..1156)
                     /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(953..972)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1501..2160)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2075..2094
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2161..2263)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2408..2639)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3237..3255)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3247..3294)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(3380..3968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3460..3479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4176..4193)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      join(4346..4348,1..84)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
       61 aacgctctcc tgagtaggac aaatccgccg ggagaacaac aacaattgca ttcattttat
      121 gtttcaggtt cagggggagg tgtgggaggt tttttaacgc tagcatggat gttttcccag
      181 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt
      241 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
      301 actttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
      361 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
      421 tgaatcaact acttagatgg tattagtgac ctgtagtcga ccgacagcct tccaaatgtt
      481 cttcgggtga tgctgccaac ttagtcgacc gacagccttc caaatgttct tctcaaacgg
      541 aatcgtcgta tccagcctac tcgctattgt cctcaatgcc gtattaaatc ataaaaagaa
      601 ataagaaaaa gaggtgcgag cctctttttt gtgtgacaaa ataaaaacat ctacctattc
      661 atatacgcta gtgtcatagt cctgaaaatc atctgcatca agaacaattt cacaactctt
      721 atacttttct cttacaagtc gttcggcttc atctggattt tcagcctcta tacttactaa
      781 acgtgataaa gtttctgtaa tttctactgt atcgacctgc agactggctg tgtataaggg
      841 agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc
      901 ggtggctgag atcagccact tcttccccga taacggagac cggcacactg gccatatcgg
      961 tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacgggaga
     1021 ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt
     1081 caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt
     1141 aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag caaaagagcc gttcatttca
     1201 ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc
     1261 agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt gacatcatat
     1321 atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag
     1381 catacctctt tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa
     1441 atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc cacgcggcgt
     1501 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1561 atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1621 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     1681 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     1741 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     1801 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1861 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1921 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg
     1981 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     2041 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     2101 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     2161 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     2221 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     2281 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa
     2341 gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt
     2401 agtcgactac aggtcactaa taccatctaa gtagttgatt catagtgact ggatatgttg
     2461 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     2521 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gttggcatta taagaaagca
     2581 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2641 ccatccagct gataagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgg
     2701 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgtacctaac
     2761 caagttcctc tttcagaggt tatttcaggc cctgcagggt taattaacaa aaaaccccgc
     2821 cggggcgggg ttttttgtta caattcttat cctcagtcct gctcctctgc cacaaagtgc
     2881 acacagttgc ctgctgggtc tctcagggca aactctctcc cccagggctg ctctccaatc
     2941 tctgtcatgg cagggccact ggcatccctg aagttggtgg agaccacctc actccactca
     3001 gcatacagct catccagtcc tctcacccac acccaagcca gggtgttgtc aggcaccacc
     3061 tggtcctgga ctgctgagat gaacagggtg acatcatctc tgaccacacc tgcaaagtca
     3121 tcctccacaa aatctctgga gaaccccaac ctgtcagtcc agaactcaac agctccagcc
     3181 acatccctgg ctgtgagcac tgggacagca ctggtcaact tggccatgat ggccctccta
     3241 tagtgagtcg tattatacta tgccgatata ctatgccgat gattaattgt caactacttt
     3301 tgaaaaattt aattaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3361 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3421 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3481 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3541 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3601 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3661 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3721 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3781 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3841 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3901 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3961 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4021 cacgttaagg gattttggtc atggctagtt aattaaccca cgtttcaaca tgcacatccc
     4081 agtaatttgg aaacattttg tttccaaaga ttcacttaac attggtttag caacatgaag
     4141 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     4201 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     4261 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     4321 gcgagagtag ggaactgcca ggcatcaa
//
