LOCUS Exported 9803 bp ds-DNA circular SYN 21-JAN-2020 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9803) AUTHORS Trial User TITLE Direct Submission JOURNAL Exported Tuesday, Jan 21, 2020 from SnapGene 5.0.7 https://www.snapgene.com FEATURES Location/Qualifiers source 1..9803 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..511 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" /note="color: #ffffff; direction: RIGHT" misc_feature 533..1240 /label=mCherry /note="color: #c0000c; direction: RIGHT" misc_feature 1283..1804 /label=SYNAPTOZIP /note="color: #84a4c0; direction: RIGHT" misc_feature 1682..1771 /label=ZIP /note="color: #ff00ff; direction: RIGHT" CDS 1772..1801 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /note="color: #cc99b2" /translation="EQKLISEEDL" misc_feature 1805..3230 /label=3'UTR VAMP2 /note="color: #339966" misc_feature 3244..3832 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" /note="color: #ffffff" CDS complement(3715..3726) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /note="color: #cc99b2 Cleavage site after base 3714" /translation="IEGR" LTR 3919..4152 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /note="color: #ffe4c4" polyA_signal 4224..4345 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" rep_origin 4364..4499 /label=SV40 ori /note="SV40 origin of replication" /note="color: #ffff00" promoter complement(4532..4550) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff; direction: LEFT" primer_bind complement(4560..4576) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" rep_origin 4721..5176 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" promoter 5205..5309 /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: RIGHT" CDS 5310..6170 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 5310 .. 5378 / #ccffcc / signal sequence 2: 5379 .. 6170 / #ccffcc Cleavage site after base 5378" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6341..6929 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" protein_bind 7217..7238 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b" promoter 7253..7283 /label=lac promoter /note="promoter for the E. coli lac operon" /note="This forward directional feature has 3 segments: 1: 7253 .. 7258 / #ffffff / -35 2: 7259 .. 7276 / #ffffff 3: 7277 .. 7283 / #ffffff / -10" protein_bind 7291..7307 /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)." /note="color: #31849b" primer_bind 7315..7331 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: RIGHT" promoter 7352..7370 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" /note="color: #ffffff; direction: RIGHT" enhancer 7471..7850 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" /note="color: #ffffff" promoter 7851..8054 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" /note="color: #ffffff; direction: RIGHT" LTR 8069..8249 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" /note="color: #ffe4c4" misc_feature 8293..8418 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" /note="color: #ffe4c4" misc_feature 8911..9144 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /note="color: #ffe4c4" misc_feature 9640..9757 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" /note="color: #ffe4c4" ORIGIN 1 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc 61 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc 121 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc 181 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac 241 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc 301 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag 361 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct 421 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct 481 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggtcgcca ccatggtgag 541 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat 601 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta 661 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg 721 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga 781 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa 841 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt 901 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa 961 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa 1021 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa 1081 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa 1141 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga 1201 gggccgccac tccaccggag gcatggacga gctgtacaag ggcaccggcg cgcctcagat 1261 ctcgagctca agcttcgaat tctcggctac cgctgccacc gtcccgcctg ccgccccggc 1321 cggcgagggt ggcccccctg cacctcctcc aaatcttacc agtaacagga gactgcagca 1381 gacccaggcc caggtggatg aggtggtgga catcatgagg gtgaatgtgg acaaggtcct 1441 ggagcgagac cagaagctat cggaactgga tgatcgcgca gatgccctcc aggcaggggc 1501 ctcccagttt gaaacaagtg cagccaagct caagcgcaaa tactggtgga aaaacctcaa 1561 gatgatgatc atcttgggag tgatttgcgc catcatcctc atcatcatca tcgtttactt 1621 cagcacttct gtccctgagg gtaagggcgt ggagccaaag acctactgct actacagcag 1681 cgctcagctg gagaaggagc tgcaggccct ggagaaggag aacgcacaac tagaatggga 1741 acttcaagcg ctcgaaaaag aactggctca ggaacaaaaa ctgatcagcg aggaggacct 1801 gtgacctgag gttcccatca cctctctctc tcaccctctg ccccctttcc tctttcctgt 1861 tcagccccaa gtcccttcat ttgcatctgc tatgcaatag cccctctcct ccttcccttg 1921 gatttaacca atccttcccc tatcttccct gtacaattcc agaccctccc caaacagagc 1981 aaaccccaca gaaacaaaca aaacacccca cctgtctagg cttccatagg ttgcatcttt 2041 gtatcccttg ggagcctcta agactggtcc tacttggtcc taagaatccc aaggttttct 2101 gggagcttcc aacatgttga ttagcatatc atttgcatac actgtctttt ttttttccct 2161 gggtttcttc ctttctgttc cattactctt cactcttctg tgtttttttt ttttctttct 2221 ttcttttcgg aagagttagt ttccattggt cctcttatca cacttttatt tgcacgacat 2281 tatctgcagg tggtggggag cctgggcttt tggggaacta ggaccccaaa gttgcctcct 2341 cccagcccct ctagtctagt ttgcttccct taccccattt tccacacctc ttgtaccctc 2401 ctctccctcc cccagctggt gtgtaagtgt cttggagttc agtgtgttat gatggaccaa 2461 taattctgcc acttggggtc tctccctaca ttcctgctcc ccagttttcg tgtggggcat 2521 tcagctgaca ttcccggggg tctccctccc tcccatctgc ctgatctgct gcctcctccc 2581 accaggagaa ctggaggctg accacaatct ggttctttga ggaggggtgg ctctagtgtg 2641 tgtgggggtc atcactgcct tggggaggag tgggacaaaa cagaaccccc cctaattcct 2701 gcctgaaatc tctggcctca tccttgctag aggttggact gaaaactttc ctccccaatc 2761 ctgggggggg gtattacccc ccatcactgc ccagctcctc tgactgcccc cctgtattca 2821 gggtgggggt actagtcact gccaatatgt gtatgggact tgctggagga tggggatcct 2881 tgtccttctc tagggctgct gagccctgag agagagagag agagagagag agagagagag 2941 agagagagag agagagagag ggaagcaagc tatttccctg ttgggtacaa tgatagagaa 3001 aggatctaat ctttaaaaat ggcacagttt taggggtctg agggtacagc cccttaacct 3061 gcctcttggg gggggtgttc cccaaactct tcccccacac acaccaggtt ttctgtgtgg 3121 aggggaacca aggagatcta aactgtggtg tgaaagggta ggagagatgc tgggggtggg 3181 ggtgcttgtg ttttagaccc ccaatattat cccagtgtcc cctgcctgaa ttctgcagtc 3241 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 3301 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 3361 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 3421 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 3481 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 3541 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 3601 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 3661 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 3721 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 3781 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt 3841 cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt 3901 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt 3961 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac 4021 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg 4081 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga 4141 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga 4201 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa 4261 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 4321 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc 4381 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 4441 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 4501 aggcttttgc gtcgagacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 4561 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 4621 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 4681 ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 4741 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 4801 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 4861 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 4921 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 4981 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 5041 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 5101 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 5161 ttaacgttta caatttccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 5221 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 5281 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 5341 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 5401 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 5461 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 5521 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 5581 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 5641 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 5701 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 5761 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 5821 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 5881 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 5941 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 6001 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 6061 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 6121 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 6181 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 6241 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 6301 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 6361 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 6421 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 6481 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 6541 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 6601 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 6661 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 6721 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 6781 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 6841 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 6901 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 6961 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 7021 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 7081 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg 7141 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca 7201 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact 7261 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa 7321 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc 7381 tggagctgca agcttggcca ttgcatacgt tgtatccata tcataatatg tacatttata 7441 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt 7501 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 7561 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 7621 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 7681 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 7741 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 7801 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt 7861 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 7921 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 7981 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 8041 atataagcag agctcgttta gtgaaccggg gtctctctgg ttagaccaga tctgagcctg 8101 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 8161 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 8221 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggacct gaaagcgaaa 8281 gggaaaccag agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc 8341 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga 8401 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt 8461 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg 8521 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa 8581 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat 8641 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa 8701 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc 8761 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa 8821 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt 8881 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc 8941 agcaggaagc actatgggcg cagcctcaat gacgctgacg gtacaggcca gacaattatt 9001 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct 9061 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag 9121 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac 9181 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca 9241 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt 9301 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa 9361 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt 9421 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt 9481 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag 9541 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc 9601 cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat 9661 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 9721 agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga 9781 gaagcttgat atcgaattcc cac //