LOCUS       Exported                6462 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pMVP destination vector, empty PiggyBac transposon vector backbone 
            w/ Hygro selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    LC001: pMVP/PB/Hygro-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6462)
  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 6462)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6462
                     /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     1921..1938
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1944..1961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1945..1961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1987..2049
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    2263..2387
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2412..2442
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2496..3155
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2562..2581)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3497..3802
                     /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     3681..3700
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3843..3967)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3982..3998)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3983..3999)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        4019..4335
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4186..4321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4248..4267
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4383..4430
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4449..5486
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5465..5473
                     /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(5526..5545)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5570..5618
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(5834..5855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   6139..6173
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     6353..6372
                     /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 ctcacatgct gtacagctta agtcatgttc tttcctgcgt tatcccctga ttctgtggat
     1861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1921 agcgagtcag tgagcgagga agctgtaaaa cgacggccag tagctccacc gccgcagcta
     1981 gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat
     2041 tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt
     2101 aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg
     2161 tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact
     2221 tttatgacta gtggatcccc cgggctgcag gaattcgata tcacaagttt gtacaaaaaa
     2281 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     2341 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gccgcattag
     2401 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
     2461 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     2521 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     2581 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     2641 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     2701 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     2761 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     2821 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     2881 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     2941 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     3001 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     3061 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     3121 aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag
     3181 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     3241 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt
     3301 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     3361 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     3421 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     3481 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     3541 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     3601 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     3661 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     3721 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     3781 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg
     3841 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     3901 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca
     3961 aagtggtgat atcaagctta tcgataccgt cgacctcgac atggtcatag ctgttgacgg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4141 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4201 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4261 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4321 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     4381 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     4441 actaaaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga
     4501 agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc cgagccttca
     4561 gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct gatggcttct
     4621 acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc cccgaagtgt
     4681 tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc gcccaaggcg
     4741 ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct gtcgccgaag
     4801 ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc ccattcgggc
     4861 cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt gctgatcccc
     4921 atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta gctcaagccc
     4981 tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc gtccatgccg
     5041 acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta atcgactggt
     5101 ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc tggcggccct
     5161 ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc gagctggccg
     5221 gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc taccagagcc
     5281 tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc
     5341 gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga
     5401 ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt acacgaccgc
     5461 gcgctaagga ggtaggtcga gtttaacgta ccggttagta atgagtttaa acgggggagg
     5521 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5581 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5641 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5701 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5761 cggggcggca ggccctgcca tagcagatcc ggttaaatca aaagaataga ccgagatagg
     5821 gttgagtgtt gttccagttt ggaacaagag tccacgctgt ccctgatatc tataacaaga
     5881 aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata taattatcgt
     5941 atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga ctcacgcggt
     6001 cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca cgggagctcc
     6061 aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt agaaagagag
     6121 agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta gggttaatct
     6181 agctgcatca agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6241 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6301 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6361 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6421 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ag
//
