LOCUS       Exported                5900 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pMVP destination vector, empty expression vector backbone w/ Hygro 
            selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    KJ801: pMVP/Hygro-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 5900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5900
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1925..1942
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1926..1942
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1982
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1982
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2008..2132
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2157..2187
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2241..2900
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2307..2326)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3242..3547
                     /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     3426..3445
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3588..3712)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3727..3743)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3728..3744)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        3764..4080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3931..4066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3993..4012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4128..4175
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4194..5231
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5210..5218
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(5271..5290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5315..5363
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      complement(5539..5897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5579..5600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5791..5810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagctgtaaa acgacggcca gtagctccac cgcggtggcg gccgctctag aactagtgga
     1981 tcccccgggc tgcaggaatt cgatatcaca agtttgtaca aaaaagctga acgagaaacg
     2041 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     2101 tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta
     2161 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt
     2221 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     2281 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     2341 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     2401 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     2461 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     2521 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     2581 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     2641 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     2701 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     2761 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     2821 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     2881 agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg
     2941 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     3001 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     3061 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     3121 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     3181 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga
     3241 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     3301 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     3361 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     3421 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     3481 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     3541 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga
     3601 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
     3661 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gtgatatcaa
     3721 gcttatcgat accgtcgacc tcgacatggt catagctgtt gacggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4081 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     4141 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaaga
     4201 agcccgaact caccgctacc agcgttgaaa aatttctcat cgagaagttc gacagtgtga
     4261 gcgacctgat gcagttgtcg gagggcgaag agagccgagc cttcagcttc gatgtcggcg
     4321 gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg cttctacaaa gaccgctacg
     4381 tgtaccgcca cttcgccagc gctgcactac ccatccccga agtgttggac atcggcgagt
     4441 tcagcgagag cctgacatac tgcatcagta gacgcgccca aggcgttact ctccaagacc
     4501 tccccgaaac agagctgcct gctgtgttac agcctgtcgc cgaagctatg gatgctattg
     4561 ccgccgccga cctcagtcaa accagcggct tcggcccatt cgggccccaa ggcatcggcc
     4621 agtacacaac ctggcgggat ttcatttgcg ccattgctga tccccatgtc taccactggc
     4681 agaccgtgat ggacgacacc gtgtccgcca gcgtagctca agccctggac gaactgatgc
     4741 tgtgggccga agactgtccc gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca
     4801 acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg
     4861 gggacagtca gtacgaggtg gccaacatct tcttctggcg gccctggctg gcttgcatgg
     4921 agcagcagac tcgctacttc gagcgccggc atcccgagct ggccggcagc cctcgtctgc
     4981 gagcctacat gctgcgcatc ggcctggatc agctctacca gagcctcgtg gacggcaact
     5041 tcgacgatgc tgcctgggct caaggccgct gcgatgccat cgtccgcagc ggggccggca
     5101 ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg
     5161 aggtgctggc cgacagcggc aaccgccggc ccagtacacg accgcgcgct aaggaggtag
     5221 gtcgagttta acgtaccggt tagtaatgag tttaaacggg ggaggctaac tgaaacacgg
     5281 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5341 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5401 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5461 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5521 tgccatagca gatccggtta aatcaaaaga atagaccgag atagggttga gtgttgttcc
     5581 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5641 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5701 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5761 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5821 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5881 gccgctacag ggcgcgtcag
//
