LOCUS       Exported                8683 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMCV-R17a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8683)
  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 8683)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8683
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1182..1976
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1236..1255)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1846..1865
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(2024..2403)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2086..2107)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2298..2317
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2428..2730)
                     /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="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2530..2549)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     2853..2875
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2867..2884
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2868..2884
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2891..2910
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2891..2909
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8427..8445)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8428..8445)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8463..8479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8463..8479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8476..8498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8487..8503
                     /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(8511..8541)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8556..8577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaagccca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt cacgtagaaa gccagtccgc
      901 agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg
      961 caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg
     1021 cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg
     1081 ggaagccctg caaagtaaac tggatggctt tctcgccgcc aaggatctga tggcgcaggg
     1141 gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     1201 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     1261 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     1321 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     1381 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     1441 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     1501 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     1561 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     1621 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     1681 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     1741 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1801 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1861 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1921 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattt
     1981 tgttaaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaacatc
     2041 ccttataaat caaaagaata gaccgcgata gggttgagtg ttgttccagt ttggaacaag
     2101 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2161 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tgcggtcgag gtgccgtaaa
     2221 gctctaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2281 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2341 gtagcggtca cgctgcgcgt aaccaccaca cccgcgcgct taatgcgccg ctacagggcg
     2401 cgtccattcg ccattcaggc ctgacattta tattccccag aacatcaggt taatggcgtt
     2461 tttgatgtca ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg
     2521 cacactggcc atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg
     2581 gtaaagttca cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat
     2641 ccgtcgcccc ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct
     2701 ctctctttta taggtgtaaa ccttaaactg ccgtacgtat aggctgcgca actgttggga
     2761 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     2821 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     2881 cagtgaattg taatacgact cactataggg cgaattgggc cctctagatg catgctcgag
     2941 cggccgccag tgtgatggat atctgcagaa ttcttctttt tcttatttcc atgttctgat
     3001 ccagggaatc tcttagattt gcctttgggg aaaagtgtaa agtataacta aatcttgcta
     3061 ttaatgtttt gggaataaaa taatcattag cagtaacaat acaaggagga aaaatctgat
     3121 gctttttatt cacatgcttc ttctctaagc ttacagctac agcaccatct agatgatctc
     3181 ttaagttatc aaggttattt attccttgcc ctggttgcag atctttattt aggctatttt
     3241 gccctttcac atcctcaaaa acaaccataa atttatccaa agcacatcct agttcaaaag
     3301 gcagtttatc agatggacag tttatattca aggccttccc ttctagcaaa tctattaagg
     3361 ctgcagcaaa gcttgttttt ccactgttaa taggcccttt aaaccaaatg tttctatact
     3421 taggtatatt ctctgttaat aattgaataa ttttctgcag cttcttttca aactcttcaa
     3481 ataagcagca gtaccaggcc acaccaccca tataatacag tagatctatt gtatctaaat
     3541 ctcttaatct ctctaggtgc ttcttaaact tcttacatag catttctgtc ctggtcattt
     3601 ccagcatctc taacctcctt ttggctagaa cagtgtctgc ggcttgttgg caaatggttt
     3661 tctgagattt agattcataa aatagcttag cattagaatg atgagcctca tgagccttgt
     3721 gaggtttgag gcgagatctg ttttcacact tttggcaagg aaatggtttt gcaaagtcta
     3781 gataatgggc taagataata aagtggtcgt ctagctcata ttcacaagca aattcagcaa
     3841 ctaaattcca attacagctg gcctcttttt ctttttcttg aaattcataa ttgagcagtg
     3901 gcttattctc ttgcagtaat ttgtaagggg gcttgcataa attattatac atttcaggca
     3961 tcttattcac tcctttacaa attaaaaagc ttatagtgca gaaggtagag caaaaattct
     4021 taatagcaga tactctatgc tttgataaag ttataaacaa taaaatacat cctaattcac
     4081 aggcatgcct gcttttaaaa tcaactttaa atttctcaat cttatcatat aactctatag
     4141 ctttatcaga agtagtataa atggcaaaac aacttactgt tttattacta tatacagcat
     4201 ggctaagata atcagaaaga tcaataggaa aatcagtagg aacaggagtt tctctgttct
     4261 tttttggctt tggtggagtg cttgtaaaac ttgctgaact agcagagctt gcagagcttc
     4321 gggacccccc aaattttcgc tttcttgaga atggaggagg ggtcttcggg gtggtgaagg
     4381 aggaggatct gtattcctca tctgtaaact gagatgacga ggcctcctcg gcagaggaag
     4441 acgggggctg ccggggcgag cttcttgagg aggggggctc ctcaggctcc tcagaggacg
     4501 agggaggctc aggggaggaa agtgattcat cgcagaagag atcctcccag gtgccatccg
     4561 ttctggaaga atttctaggt acactggttc cattgggtgt gctggattct cttcctgaat
     4621 tggtggtctc ctctctgcta ctggatccag aggatgaggt gggttcctca tggtgttcgg
     4681 gaggtatatc gggtcctctg gactgggagt ctgaagcctg ggacgctgag aaggacccat
     4741 acccagagga agagctctgg ctgtggggtg gtgagcttcc actgggggct cccctggatg
     4801 cattggagga aggctttctg gatcttgagt tggtcccgtg tggattgggc ccatattcgt
     4861 atgccttccc gaagctgaat cctcctgatc tccaccattc tttgaattta gtggtcccat
     4921 atataggggc ctcgtcaacc tagatgggaa agtacagaaa atctgtcata aataaccttt
     4981 ctttgatatt ttgccttata gacttttcca tatctaatac ttacagagga aggaagtagg
     5041 agtctagaaa aggtgcagat gcagtaagca gtagtcagtt tcttctaaag ttttttgcca
     5101 ccagtcaaaa ctttcccaag taggaggaaa tccaaaccaa agaataaagc actgatagca
     5161 aaaacactct ccccacgtca gacagttttt ttgctttaaa gtttttagac tacaatgctg
     5221 gcgagacaac ttacagctaa tacaagcgca cttagaatct ctaagttgct taagcatgca
     5281 cccaggacct ctgcaaaatc tagcattata tccactttgc atataatcct ttaaagttcc
     5341 atattcttcc caaggaaatt ttgtactgac ctcatcaaac atagagaagt cacttctgag
     5401 cttgtggata ttttgctgga atttgctcca aagggtgttc aattccatca ttataacagg
     5461 atttccccct ttatcagggt gatgctttaa gcagcttctt ttgaaagcag ctttcatcag
     5521 agggatgttg ccataacaat taggagcaat ctctaaaagc ttgcagagag cctctctttc
     5581 tttcctattt aggactaaat ccatcttgtc tatatgcaga aggagtttgc agaaagagca
     5641 gaggagcaaa tgagctacct cactaaggag tggtttttat actgcagttt cccgcccttg
     5701 ggatctgccc ttagatactg ccttttttgc taattaagcc tcttaagcct cagaggcctc
     5761 tctctttttt tccagaggcc tcggaggcta ggagccccaa gcctctgcca acttgaaaaa
     5821 aaaaagtcac ctaggcagcc aagttgtggt tacatgattg aacttttatt gctgcagggt
     5881 ttctggcatt gactcatttc ctggagaggc ggagtttgac tgataaacaa aacttttttt
     5941 ctttctgttt gggagggaga cggaagactc ttaacttttt ttcaacaagg gaggcccgga
     6001 ggcttttttt tctcttacaa agggaggagg acattaaaag agtaagtatc cttatttatt
     6061 tttcaggatg gggggcatca tcacactgct ggccaatatt ggtgaaattg ctactgaact
     6121 aagtgccacc acaggagtaa ctttggaagc tattcttaca ggagaagctt tagcagcttt
     6181 ggaagcagag atctccagtt taatgacaat tgagggtatt tctggcattg aggctttagc
     6241 tcaacttggg ttcacagctg aacagttttc aaatttctca ttagtggctt ctttggttaa
     6301 ccaaggttta acttatggct tcattctcca aactgttagt ggtataggct ctctaataac
     6361 tgtgggggtg aggttgtcac gcgagcaagt gtcacttgta aagagggatg tttcgtgggt
     6421 aggtagtaat gaggttttga ggcatgcact tatggccttt agcctagatc ctctgcagtg
     6481 ggaaaatagc ttgctgcatt ctgtggggca agatattttt aattctttat ctcctacctc
     6541 taggctgcag atacaatcaa acctagtgaa tctgatacta aatagccggt gggtctttca
     6601 gacaactgct tctcagaatc agggcctttt atcaggagag gctatattaa ttcctgaaca
     6661 tataggagga actctgcagc agcaaactcc agattggctt cttcctctgg tactaggcct
     6721 tagtggatat atttctcctg aattacaagt aattgaagat ggcaccaaaa agaaaagcat
     6781 catccacctg taaaacaccc aaaaggcaat gtatacctaa gccgggatgc tgccctaatg
     6841 ttgcctcagt tccaaaactg cttgttaaag gaggagtgga agtattatct gtggttactg
     6901 gagaagatag cattacccaa attgagttgt atttgaatcc aagaatggga gttaattccc
     6961 ctgatcttcc tactacttca aactggtata cttatactta tgacctgcag ccaaagggat
     7021 catctccaga tcagcccatc aaggaaaatt tgccagctta cagtgtggca agagtgtctc
     7081 tgccaatgct aaatgaggat attacctgtg acacattgca gatgtgggag gcaatatctg
     7141 ttaaaacaga agtagttgga ataagttctt taattaatgt tcattattgg gacatgaaaa
     7201 gagttcatga ttatggtgct ggtattcctg tgtcaggggt aaattaccat atgtttgcca
     7261 ttgggggaga acctctggat ttgcaaggcc tagttttaga ttaccagact gagtatccaa
     7321 aaactacaaa tggtgggcct attacaattg aaactgtatt gggaagaaaa atgacaccta
     7381 aaaatcaggg cctagatcca caagctaaag caaaattaga taaagatgga aattatccta
     7441 tagaagtatg gtgtcctgat ccttctaaaa atgaaaacag tagatactat gggtctattc
     7501 agacaggctc tcagactcct acagttcttc aatttagtaa tactctaact actgtccttt
     7561 tagatgagaa tggagtgggc cctctatgca aaggagatgg cctatttatt agctgtgcag
     7621 acatagtggg gtttctgttt aaaaccagtg gaaaaatggc tcttcatggg ttgcctagat
     7681 attttaatgt tactttgaga aaaagatggg tgaaaaaccc ctacccagta gttaatttaa
     7741 taaactcact cttcagcaac ttaatgccaa aagtgtcagg ccaacctatg gaaggaaaag
     7801 ataatcaggt agaagaggtt agaatatatg aggggtcaga acaattacct ggtgatcctg
     7861 atattgtcag atttttagat aaatttgggc aggagaaaac tgtttaccca aagccctctg
     7921 ttgccccagc agcagtaaca ttccaaagta atcagcagga taagggcaag gcgccactga
     7981 aaggacctca aaaggcctct caaaaagaaa gccaaacaca agaattatga gaattatttc
     8041 atgcattcct attcagttaa gtaggcccca gaaaaacaaa cacaggaaat atgaagcaga
     8101 tgcctttatt gagaaaaagt accagaatct tgggtttctt cagtttcctc agggccctct
     8161 tcctcaataa gaatattgag cagagggtcc tgaccagctt ctacattttc tatcatttga
     8221 caaaatttac catatgatat ttcactctgt aaaatttgct tccagttttt aatttcttct
     8281 tgtaagcaag gcttaaaggt tgtatcaggc aagcaccaaa taagacaaag caataaagtg
     8341 gttccacttt gaagaattcc agcacactgg cggccgttac tagtggatcc gagctcggta
     8401 ccaagcttga tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca
     8461 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     8521 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     8581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     8641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gct
//
