LOCUS Exported 7266 bp ds-DNA circular SYN 05-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCDH-EF1-MCS-T2A-copGFP (CD521A-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7266)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7266
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 234..414
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 458..583
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1076..1309
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1804..1921
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1966..2177
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2190..2458
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 2524..2577
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/note="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 2608..3270
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/note="CopGFP"
/translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
misc_feature 3361..3949
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(3832..3843)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 4023..4256
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4328..4449
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4468..4603
/note="SV40 ori"
/note="SV40 origin of replication"
primer_bind complement(4641..4657)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4665..4681
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(4689..4719)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4734..4755
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5043..5631)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5802..6662)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6663..6767)
/gene="bla"
/note="AmpR promoter"
primer_bind 7241..7257
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
1921 tatcgatact agtaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
1981 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
2041 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
2101 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
2161 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
2221 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
2281 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
2341 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
2401 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
2461 agctgtgacc ggcgcctact ctagagctag cgaattcgaa tttaaatcgg atccgcggcc
2521 gctgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggcccttcc
2581 ggaatggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc
2641 accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc caagcagggc
2701 cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg
2761 agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac
2821 cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag
2881 gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc
2941 gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc
3001 atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt gctggtgggc
3061 agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt ggtggacagc
3121 cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc
3181 gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt ggagtaccag
3241 cacgccttca agacccccat cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc
3301 gtggacggca ccgccggacc cggctccacc ggatctcgct agagctgaat ctaagtcgac
3361 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
3421 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
3481 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
3541 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
3601 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
3661 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
3721 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
3781 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
3841 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
3901 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt
3961 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
4021 actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc
4081 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
4141 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
4201 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
4261 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
4321 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
4381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
4441 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat
4501 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
4561 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac
4621 ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
4681 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
4741 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
4801 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
4861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
4921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
4981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
5041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
5101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
5161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
5221 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
5281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
5341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
5401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
5461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
5521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
5581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
5641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
5701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
5761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
5821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
5881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
5941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
6001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
6061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
6121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
6181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
6241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
6301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
6361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
6421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
6481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
6541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
6601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
6661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
6721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
6781 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
6841 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
6901 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
6961 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
7021 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
7081 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
7141 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
7201 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
7261 aagctg
//