LOCUS Exported 8787 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 23
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8787)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8787
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/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 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 2793..2873
/note="MCS"
/note="multiple cloning site"
CDS 2875..3594
/codon_start=1
/product="Aequorea coerulescens GFP"
/note="AcGFP1"
/note="mammalian codon-optimized"
/translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
GFVTAAAITHGMDELYK"
promoter 3622..4121
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 4142..4741
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4755..5343
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5226..5237)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5550..6183
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6312..6328)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6336..6352
/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(6360..6390)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6405..6426
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6714..7302)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7473..8333)
/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(8334..8438)
/gene="bla"
/note="AmpR promoter"
polyA_signal 8486..8620
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcatggtg
2881 agcaagggcg ccgagctgtt caccggcatc gtgcccatcc tgatcgagct gaatggcgat
2941 gtgaatggcc acaagttcag cgtgagcggc gagggcgagg gcgatgccac ctacggcaag
3001 ctgaccctga agttcatctg caccaccggc aagctgcctg tgccctggcc caccctggtg
3061 accaccctga gctacggcgt gcagtgcttc tcacgctacc ccgatcacat gaagcagcac
3121 gacttcttca agagcgccat gcctgagggc tacatccagg agcgcaccat cttcttcgag
3181 gatgacggca actacaagtc gcgcgccgag gtgaagttcg agggcgatac cctggtgaat
3241 cgcatcgagc tgaccggcac cgatttcaag gaggatggca acatcctggg caataagatg
3301 gagtacaact acaacgccca caatgtgtac atcatgaccg acaaggccaa gaatggcatc
3361 aaggtgaact tcaagatccg ccacaacatc gaggatggca gcgtgcagct ggccgaccac
3421 taccagcaga atacccccat cggcgatggc cctgtgctgc tgcccgataa ccactacctg
3481 tccacccaga gcgccctgtc caaggacccc aacgagaagc gcgatcacat gatctacttc
3541 ggcttcgtga ccgccgccgc catcacccac ggcatggatg agctgtacaa gtgagcggcc
3601 gcgactctag ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
3661 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
3721 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
3781 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
3841 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
3901 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
3961 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
4021 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
4081 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac
4141 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
4201 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
4261 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
4321 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
4381 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
4441 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
4501 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
4561 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
4621 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
4681 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
4741 accgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
4801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
4861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
4921 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
4981 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
5041 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
5101 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
5161 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
5221 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
5281 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
5341 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
5401 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
5461 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
5521 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
5581 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
5641 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
5701 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
5761 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
5821 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
5881 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
5941 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
6001 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
6061 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
6121 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
6181 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
6241 cagagagtga gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg
6301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
6361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
6421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
6481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
6541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
6601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
6661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
6721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
6781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
6841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
6901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
6961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
7021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
7081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
7141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
7201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
8461 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
8521 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
8581 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
8641 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
8701 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
8761 ttaaatatgt actacaaact tagtagt
//