LOCUS Exported 8825 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 22
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8825)
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..8825
/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)"
promoter 2338..3519
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2568..3510
/note="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
misc_feature 3575..4148
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4181..4780
/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 4794..5382
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5265..5276)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5589..6222
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6350..6366)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6374..6390
/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(6398..6428)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6443..6464
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6752..7340)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7511..8371)
/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(8372..8476)
/gene="bla"
/note="AmpR promoter"
polyA_signal 8524..8658
/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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc
3541 ctcgagacta gttctagagc ggccgcggat cccgcccctc tccctccccc ccccctaacg
3601 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
3661 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
3721 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
3781 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
3841 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
3901 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
3961 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
4021 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
4081 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
4141 gatgataagc ttgccacaac ccacaaggag acgaccttcc atgaccgagt acaagcccac
4201 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
4261 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
4321 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
4381 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
4441 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
4501 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
4561 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
4621 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
4681 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
4741 accgcgcacc tggtgcatga cccgcaagcc cggtgcctag acgcgtctgg aacaatcaac
4801 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
4861 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
4921 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
4981 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
5041 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
5101 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
5161 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
5221 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
5281 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
5341 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
5401 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
5461 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
5521 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
5581 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
5641 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
5701 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
5761 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
5821 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
5881 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
5941 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
6001 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
6061 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
6121 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
6181 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
6241 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
6301 attgctagcg tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
6361 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
6421 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
6481 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
6541 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
6601 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
6661 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
6721 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
6781 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
6841 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
6901 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
6961 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
7021 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
7081 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
7141 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
7201 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
7261 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
7321 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
7381 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
7441 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
7501 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
7561 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
7621 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
7681 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
7741 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
7801 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
7861 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
7921 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
7981 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
8041 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
8101 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
8161 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
8221 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
8281 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
8341 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
8401 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
8461 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
8521 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
8581 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
8641 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
8701 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
8761 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
8821 gtagt
//