LOCUS Exported 8931 bp ds-DNA circular SYN 23-JUN-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRRLSIN.cPPT.PGK-GFP.WPRE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8931)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Friday, June 23, 2017 from SnapGene Viewer 3.3.4
http://www.maiolingbio.com
FEATURES Location/Qualifiers
source 1..8931
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..227
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 228..408
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 455..580
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1073..1306
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 3345..3462
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 3508..4018
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 4040..4759
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 4775..5363
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5246..5257)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5450..5683
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5755..5876
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 5916..6051
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(6072..6090)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6100..6116)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6258..6713
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6739..6843
/gene="bla"
/label=AmpR promoter
CDS 6844..7704
/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"
rep_origin 7875..8463
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 8751..8772
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8787..8817
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8825..8841
/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 8849..8865
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 8886..8904
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacctgtct cttgatcaga
1441 tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca
1501 gggcttccca accttcccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca
1561 taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt
1621 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
1681 ccgtttctgc ggactggctt tctacgtgtt ccgcttcctt tagcagccct tgcgccctga
1741 gtgcttgcgg cagcgtgaag ctttctctga gctgtaacag cctgaccgca acaaacgaga
1801 ggatcgagac catccgctcc agattatccg gctcctccat gcgttgcctc tcggctcctg
1861 ctccggtttt ccatgcctta tggaactcct cgatccgcca gcgatgggta taaatgtcga
1921 tgacgcgcaa ggcttgggct agcgactcga ccggttcgct ggtcagcaac aaccatttca
1981 acggggtctc acccttgggc gggttaatct cctcggccag caccgcgttg agcgtgatat
2041 tcccctgttt tagcgtgatg cgcccactgc gcaggctcaa gctcgccttg cgggctggtc
2101 gatttttacg tttaccgcgt ttatccacca cgcccttttg cggaatgctg atctgatagc
2161 cacccaactc cggttggttc ttcagatggt cgtacagata caacccagac tctacgtcct
2221 tgcgtgggtg cttggagcgc accacgaagc gctcgttatg cgccagtttg tcctgcagat
2281 aagcatgaat atcggcttcg cggtcacaga ccgcaatcac gttgctcatc atgctgccca
2341 tgcgtaaccg gctagttgcg gccgctgcca gccatttgcc actctccttt tcatccgcat
2401 cggcagggtc atccgggcgc atccaccact cctgatgcag taatcctacg gtgcggaatg
2461 tggtggcctc gagcaagaga acggagtgaa cccaccatcc gcgggattta tcctgaatag
2521 agcccagctt gccaagctct tcggcgacct ggtggcgata actcaaagag gtggtgtcct
2581 caatggccag cagttcggga aactcctgag ccaacttgac tgtttgcatg gcgccagcct
2641 ttctgatcgc ctcggcagaa acgttgggat tgcggataaa tcggtaagcg ccttcctgca
2701 tggcttcact accctctgat gagatggtta ttgatttacc agaatatttt gccaattggg
2761 cggcgacgtt aaccaagcgg gcagtacggc gaggatcacc cagcgccgcc gaagagaaca
2821 cagatttagc ccagtcggcc gcacgatgaa gagcagaagt tatcatgaac gttaccatgt
2881 taggaggtca catggaagat cagatcctgg aaaacgggaa aggttccgtt caggacgcta
2941 cttgtgtata agagtcagga gtgggacaga gaaattaaca attacacaag cttaatacac
3001 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
3061 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
3121 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
3181 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
3241 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
3301 agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa aagaaaaggg
3361 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
3421 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttatcgatca cgagactagc
3481 ctcgagaagc ttgatatcga attccacggg gttggggttg cgccttttcc aaggcagccc
3541 tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca gcggcgccga
3601 ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct tcgccgctac
3661 ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag gttccttgcg
3721 gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta ccctcgcaga
3781 cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg
3841 ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg
3901 tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc
3961 ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg
4021 ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
4081 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
4141 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
4201 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
4261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
4321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
4381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
4441 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
4501 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
4561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
4621 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
4681 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
4741 ggacgagctg tacaagtaaa gcggccgcgt cgacaatcaa cctctggatt acaaaatttg
4801 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
4861 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
4921 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
4981 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
5041 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
5101 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
5161 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
5221 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
5281 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
5341 ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacctttaag accaatgact
5401 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
5461 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
5521 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
5581 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
5641 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
5701 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
5761 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
5821 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
5881 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
5941 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
6001 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
6061 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
6121 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
6181 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
6241 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
6301 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
6361 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
6421 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
6481 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
6541 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
6601 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
6661 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
6721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
6781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
6841 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
6901 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
6961 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
7021 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
7081 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
7141 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
7201 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
7261 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
7321 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
7381 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
7441 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
7501 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
7561 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
7621 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
7681 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
7741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
7801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
7861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
7921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
7981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
8041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
8101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
8161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
8221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
8281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
8341 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
8401 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
8461 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
8521 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
8581 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
8641 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
8701 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
8761 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
8821 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
8881 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta a
//