LOCUS Exported 9227 bp ds-DNA circular SYN 03-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..9227
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/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 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
polyA_signal 2187..2321
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 2496..2527
/label=MCS
promoter complement(2528..2895)
/label=TRE3GS promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, modified to eliminate
binding sites for endogenous mammalian transcription
factors"
promoter 2912..3422
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 3441..4187
/codon_start=1
/product="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
/label=Tet-On 3G
/translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
promoter 4198..4527
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4378..4513
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4536..5135
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 5149..5737
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5945..6578
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
rep_origin complement(7109..7694)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7865..8725)
/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"
promoter complement(8726..8830)
/gene="bla"
/label=AmpR promoter
polyA_signal 8878..9012
/label=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 actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgacttaact tgtttattgc agcttataat ggttacaaat
2221 aaggcaatag catcacaaat ttcacaaata aggcattttt ttcactgcat tctagttttg
2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctcaaatccc tcggaagctg
2341 cgcctgtctt aggttggagt gatacatttt tatcactttt acccgtcttt ggattaggca
2401 gtagctctga cggccctcct gtcttaggtt agtgaaaaat gtcactctct tacccgtcat
2461 tggctgtcca gcttagctcg caggggaggt ggtctggatc cgccggcacc ggtgtatacg
2521 ggaattcttt acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg
2581 gaattgaagt ttactcaaaa aatcagcact cttttatagg cgccctggtt tacataagca
2641 aagcttatac gttctctatc actgataggg agtaaactgg atatacgttc tctatcactg
2701 atagggagta aactgtagat acgttctcta tcactgatag ggagtaaact ggtcatacgt
2761 tctctatcac tgatagggag taaactcctt atacgttctc tatcactgat agggagtaaa
2821 gtctgcatac gttctctatc actgataggg agtaaactct tcatacgttc tctatcactg
2881 atagggagta aactcgaggt gataattcca cggggttggg gttgcgcctt ttccaaggca
2941 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
3001 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
3061 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
3121 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
3181 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
3241 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
3301 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
3361 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
3421 agggggatca tcgaattacc atgtctagac tggacaagag caaagtcata aactctgctc
3481 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
3541 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
3601 ccctgccaat cgagatgctg gacaggcatc atacccactc ctgccccctg gaaggcgagt
3661 catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc
3721 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
3781 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
3841 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
3901 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg aaacaagcaa
3961 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
4021 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg
4081 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
4141 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaacg cgcgaatgtg
4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
4441 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
4501 ttttttggag gcctaggctt ttgcaaaacg cgaccatgac cgagtacaag cccacggtgc
4561 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
4621 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
4681 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
4741 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
4801 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
4861 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
4921 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
4981 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
5041 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
5101 gcacctggtg catgacccgc aagcccggtg cctgaacgcg tctggaacaa tcaacctctg
5161 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
5221 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
5281 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
5341 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
5401 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
5461 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
5521 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
5581 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
5641 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
5701 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
5761 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
5821 cctggctaga agcacaagag gaggaggagg tgggtttttc cagtcacacc tcaggtacct
5881 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg
5941 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
6001 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat
6061 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag
6121 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac
6181 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc
6241 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt
6301 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc
6361 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat
6421 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
6481 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
6541 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
6601 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg
6661 ctagcgtttt accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
6721 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
6781 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
6841 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
6901 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
6961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
7021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
7081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
7141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
7201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
7261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
7321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
7381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
7441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
7501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
7561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
7621 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
7681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
7741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
7801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
7861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
7921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
7981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
8041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
8101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
8161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
8221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
8281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
8341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
8401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
8461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
8521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
8581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
8641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
8701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
8761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
8821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac
8881 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
8941 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
9001 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
9061 taccctatta ccactgccaa ttacctagtg gtttcattta ctctaaacct gtgattcctc
9121 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
9181 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//