LOCUS       Exported                8481 bp ds-DNA     circular SYN 21-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   VB221224-1046ybg
VERSION     .
KEYWORDS    VB221224-1046ybg.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8481)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8481
                     /organism="synthetic DNA construct"
     promoter        1..229
                     /label=RSV  promoter
                     /note="/application_notes=Strong promoter; drives 
                     transcription
                     of viral RNA in packaging cells."
                     /note="/description=Rous sarcoma virus enhancer/promoter"
                     /note="/full_name=Rous sarcoma virus (RSV) 
                     enhancer/promoter"
                     /note="/official_designation=RSV"
                     /note="/uuid=3f2ddca7f0cb4015b6abcec8a25c834e"
     LTR             230..410
                     /label=5' LTR-Delta-U3
                     /note="/application_notes=Allows transcription of viral RNA
                     and
                     its packaging into virus."
                     /note="/description=Truncated HIV-1 5' long terminal 
                     repeat"
                     /note="/full_name=HIV-1 truncated 5' LTR"
                     /note="/official_designation=Delta5' LTR"
                     /note="/uuid=44156a3083604321be44066133e8ecb9"
     misc_feature    521..565
                     /label=Psi
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Allows packaging of viral RNA 
                     into
                     virus."
                     /note="/description=HIV-1 packaging signal"
                     /note="/full_name=HIV-1 psi packaging signal"
                     /note="/official_designation=Psi"
                     /note="/uuid=284899413ff943a99f8210a11ad8c54e"
     misc_feature    1075..1308
                     /label=RRE
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Rev protein binding site that 
                     allows
                     Rev-dependent nuclear export of viral RNA during viral 
                     packaging."
                     /note="/description=HIV-1 Rev response element"
                     /note="/full_name=HIV-1 Rev response element"
                     /note="/official_designation=RRE"
                     /note="/uuid=df07099e49674cf3aba3661b54e28bd7"
     misc_feature    1803..1920
                     /label=cPPT
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Facilitates the nuclear import of
                     HIV-1
                     cDNA through a central DNA flap."
                     /note="/description=Central polypurine tract"
                     /note="/full_name=Central polypurine tract"
                     /note="/official_designation=cPPT"
                     /note="/uuid=f7cb29985f9f41a7a5924d3462266f42"
     promoter        1959..2190
                     /label=EFS
                     /note="/application_notes=Medium-strength promoter."
                     /note="/description=Human eukaryotic translation elongation
                     factor 1 alpha1 short form"
                     /note="/full_name=Human eukaryotic translation elongation 
                     factor
                     1 alpha1 short promoter"
                     /note="/official_designation=EFS"
                     /note="/uuid=3ff4e4bac3a04624bac899ae4a7baa8d"
     misc_feature    2215..2220
                     /label=Kozak
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Facilitates translation 
                     initiation of
                     ATG start codon downstream of the Kozak sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
     misc_feature    2221..3153
                     /label={mCherry2-NLS-PEST}
                     /label=nonstandard type:  ORF
                     /note="/application_notes="
                     /note="/official_designation={mCherry2-NLS-PEST}"
                     /note="/uuid=9ba3d5c67c084fab85a2264a38dd0760"
     primer_bind     complement(2465..2481)
                     /label=702
     primer_bind     2517..2538
                     /label=new
     primer_bind     complement(2577..2596)
                     /label=701
     primer_bind     complement(2583..2606)
                     /label=new_rev
     primer_bind     2704..2723
                     /label=704
     misc_feature    3192..3789
                     /label=WPRE
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Enhances virus stability in 
                     packaging
                     cells, leading to higher titer of packaged virus; enhances 
                     higher expression of transgenes."
                     /note="/description=Woodchuck hepatitis virus 
                     posttranscriptional
                     regulatory element"
                     /note="/full_name=Woodchuck hepatitis virus 
                     posttranscriptional
                     regulatory element"
                     /note="/official_designation=WPRE"
                     /note="/uuid=0aedff8308474b369c622433538d7836"
     primer_bind     complement(3249..3269)
                     /label=56
     primer_bind     3647..3663
                     /label=651
     promoter        3808..4318
                     /label=mPGK promoter
                     /note="/application_notes=Medium-strength promoter."
                     /note="/description=Mouse phosphoglycerate kinase 1 
                     promoter"
                     /note="/full_name=Mouse phosphoglycerate kinase 1 promoter"
                     /note="/official_designation=mPGK promoter"
                     /note="/uuid=bce93795b5fd4173b8b9abc832edad80"
     primer_bind     complement(3885..3904)
                     /label=541
     primer_bind     4247..4266
                     /label=175
     misc_feature    4331..4930
                     /label=Puro
                     /label=nonstandard type:  ORF
                     /note="/application_notes=Allows cells to be resistant to
                     puromycin."
                     /note="/description=Puromycin resistance gene"
                     /note="/full_name=Puromycin resistance gene"
                     /note="/official_designation=Puro"
                     /note="/uuid=39d7413208494d9291d10b21b13ca44c"
     primer_bind     complement(4466..4484)
                     /label=348
     primer_bind     complement(4646..4664)
                     /label=350
     LTR             5001..5235
                     /label=3' LTR-Delta-U3
                     /note="/application_notes=Allows packaging of viral RNA 
                     into
                     virus; self-inactivates the 5' LTR by a copying mechanism 
                     during viral genome integration; contains polyadenylation 
                     signal for transcription termination."
                     /note="/description=Truncated HIV-1 3' long terminal 
                     repeat"
                     /note="/full_name=HIV-1 truncated 3' LTR"
                     /note="/official_designation=DeltaU3/3' LTR"
                     /note="/uuid=c958755414c34372b909e20e81ebcef6"
     polyA_signal    5308..5442
                     /label=SV40 early pA
                     /note="/application_notes=Allows transcription termination 
                     and
                     polyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/description=Simian virus 40 early polyadenylation 
                     signal"
                     /note="/full_name=SV40 early polyadenation signal"
                     /note="/official_designation=SV40 early pA"
                     /note="/uuid=2fcb13a62e704d8095b188d9625a6a94"
     primer_bind     complement(5398..5429)
                     /label=580
     primer_bind     complement(6019..6043)
                     /label=474
     misc_feature    6396..7256
                     /label=AmpiciIIin
                     /label=nonstandard type:  ORF
                     /note="/application_notes=Allows E. coli to be resistant to
                     ampiciIIin."
                     /note="/description=AmpiciIIin resistance gene"
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=6a7994dc58024d80b36e4a85ad2e1d99"
     rep_origin      7427..8015
                     /direction=RIGHT
                     /label=pUC ori
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.
                     coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=64a8cb3def9f49deb6752795c2d893a8"
     primer_bind     complement(8056..8075)
                     /label=473
     primer_bind     complement(8056..8075)
                     /label=334
     primer_bind     complement(8352..8371)
                     /label=351
     primer_bind     complement(8367..8386)
                     /label=353
     primer_bind     complement(8414..8433)
                     /label=355
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga 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 ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttggg ctccggtgcc cgtcagtggg
     1981 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2041 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2101 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2161 ttcgcaacgg gtttgccgcc agaacacagg caagtttgta caaaaaagca ggctgccacc
     2221 atggtgagca agggcgagga ggacaacatg gccatcatca aggagttcat gaggttcaag
     2281 gtgcacatgg agggcagcgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     2341 aggccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     2401 ttcgcctggg acatcctgag cccccagttc atgtacggca gcaaggccta cgtgaagcac
     2461 cccgccgaca tccccgacta cctgaagctg agcttccccg agggcttcaa ctgggagagg
     2521 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggacagcag cctgcaggac
     2581 ggcgagttca tctacaaggt gaagctgagg ggcaccaact tccccagcga cggccccgtg
     2641 atgcagtgca ggaccatggg ctgggaggcc agcaccgaga ggatgtaccc cgaggacggc
     2701 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgcc
     2761 gaggtgaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtg
     2821 gacatcaagc tggacatcct gagccacaac gaggactaca ccatcgtgga gcagtacgag
     2881 agggccgagg gcaggcacag caccggcggc atggacgagc tgtacaagct cagatctcga
     2941 gctgatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa
     3001 aagaagagaa aggtaggatc caccggatct agccatggct tcccgccgga ggtggaggag
     3061 caggatgatg gcacgctgcc catgtcttgt gcccaggaga gcgggatgga ccgtcaccct
     3121 gcagcctgtg cttctgctag gatcaatgtg tagacccagc tttcttgtac aaagtggtga
     3181 taatcgaatt ccgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3241 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3301 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3361 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3421 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3481 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3541 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     3601 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3661 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3721 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3781 ccgcatcggg aattcccgcg gttcgaattc taccgggtag gggaggcgct tttcccaagg
     3841 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     3901 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     3961 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     4021 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     4081 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     4141 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     4201 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     4261 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctca
     4321 cgtggccacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     4381 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     4441 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     4501 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     4561 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4621 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4681 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4741 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4801 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4861 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4921 cggtgcctga ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     4981 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     5041 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5101 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5161 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5221 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     5281 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     5341 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5401 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     5461 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5521 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5581 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     5641 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5701 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5761 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     5821 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5881 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5941 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6001 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6061 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6121 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6181 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6241 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6361 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6421 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6481 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6541 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6601 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6661 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6721 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6781 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6841 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6901 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6961 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7021 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7081 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7141 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7201 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7261 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7321 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7381 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7441 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7501 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7561 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7621 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7681 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7741 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7801 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagagaga aaggcggaca
     7861 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7921 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7981 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8041 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8101 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8161 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     8221 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     8281 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     8341 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     8401 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     8461 aaaagctgga gctgcaagct t
//
