LOCUS       Exported                8555 bp ds-DNA     circular SYN 04-APR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8555)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8555
                     /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..2264
                     /label={HKGT1.1_XbaI}
                     /note="/application_notes="
                     /note="/official_designation={HKGT1.1_XbaI}"
                     /note="/uuid=688596074fea42cda59a903180239af1"
     misc_feature    2289..2294
                     /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    2295..3227
                     /label={mCherry2-NLS-PEST}
                     /label=nonstandard type:  ORF
                     /note="/application_notes="
                     /note="/official_designation={mCherry2-NLS-PEST}"
                     /note="/uuid=9ba3d5c67c084fab85a2264a38dd0760"
     primer_bind     2389..2403
                     /label=1630
                     /note="1630"
     primer_bind     complement(2651..2670)
                     /label=701
                     /note="701"
     primer_bind     2778..2797
                     /label=704
                     /note="704"
     primer_bind     complement(2983..3002)
                     /label=876
                     /note="876"
     primer_bind     complement(2983..3002)
                     /label=1116
                     /note="1116"
     misc_feature    3266..3863
                     /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(3323..3343)
                     /label=56
                     /note="56"
     primer_bind     3721..3737
                     /label=651
                     /note="651"
     promoter        3882..4392
                     /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(3959..3978)
                     /label=541
                     /note="541"
     primer_bind     4321..4340
                     /label=175
                     /note="175"
     misc_feature    4405..5004
                     /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     4405..4424
                     /label=1560
                     /note="1560"
     primer_bind     4405..4424
                     /label=1586
                     /note="1586"
     primer_bind     complement(4540..4558)
                     /label=348
                     /note="348"
     primer_bind     complement(4542..4560)
                     /label=1587
                     /note="1587"
     primer_bind     complement(4720..4738)
                     /label=350
                     /note="350"
     LTR             5075..5309
                     /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    5382..5516
                     /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(5457..5480)
                     /label=758
                     /note="758"
     primer_bind     complement(6093..6117)
                     /label=474
                     /note="474"
     misc_feature    6470..7330
                     /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      7501..8089
                     /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(8130..8149)
                     /label=473
                     /note="473"
     primer_bind     complement(8389..8407)
                     /label=1002
                     /note="1002"
     primer_bind     complement(8426..8445)
                     /label=351
                     /note="351"
     primer_bind     complement(8441..8460)
                     /label=353
                     /note="353"
     primer_bind     complement(8488..8507)
                     /label=355
                     /note="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 aaaagttgtc tagagcgcga ctttttttcc
     1981 cggcggcccc gcggctctgc ccctcgtgcg gtcgcgttcc cggcggcgcg gggggcgtgg
     2041 cctgggcgca gcatctataa aggcgggcgg cggcagaggc gccattttgc gaacggcgag
     2101 cagcggcggc ggcgaagcgc ccgccggggg cggggagggg gcggggccat ccgcgtgagg
     2161 cgaccctgtt ggtccggagg ggcggggcga ggaggaggac ccgcttgggc ggttcggctg
     2221 cccacagtaa ccgctgggtg gacctggcca gcgctccgaa ccttcaagtt tgtacaaaaa
     2281 agcaggctgc caccatggtg agcaagggcg aggaggacaa catggccatc atcaaggagt
     2341 tcatgaggtt caaggtgcac atggagggca gcgtgaacgg ccacgagttc gagatcgagg
     2401 gcgagggcga gggcaggccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     2461 gcggccccct gcccttcgcc tgggacatcc tgagccccca gttcatgtac ggcagcaagg
     2521 cctacgtgaa gcaccccgcc gacatccccg actacctgaa gctgagcttc cccgagggct
     2581 tcaactggga gagggtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggaca
     2641 gcagcctgca ggacggcgag ttcatctaca aggtgaagct gaggggcacc aacttcccca
     2701 gcgacggccc cgtgatgcag tgcaggacca tgggctggga ggccagcacc gagaggatgt
     2761 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2821 gccactacga cgccgaggtg aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2881 gcgcctacaa cgtggacatc aagctggaca tcctgagcca caacgaggac tacaccatcg
     2941 tggagcagta cgagagggcc gagggcaggc acagcaccgg cggcatggac gagctgtaca
     3001 agctcagatc tcgagctgat ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa
     3061 aggtagatcc aaaaaagaag agaaaggtag gatccaccgg atctagccat ggcttcccgc
     3121 cggaggtgga ggagcaggat gatggcacgc tgcccatgtc ttgtgcccag gagagcggga
     3181 tggaccgtca ccctgcagcc tgtgcttctg ctaggatcaa tgtgtagacc cagctttctt
     3241 gtacaaagtg gtgataatcg aattccgata atcaacctct ggattacaaa atttgtgaaa
     3301 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3361 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3421 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3481 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3541 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3601 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3661 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3721 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3781 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3841 tttgggccgc ctccccgcat cgggaattcc cgcggttcga attctaccgg gtaggggagg
     3901 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     3961 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     4021 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     4081 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
     4141 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     4201 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     4261 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     4321 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     4381 gcctttcgac ctcacgtggc caccatgacc gagtacaagc ccacggtgcg cctcgccacc
     4441 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4501 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4561 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4621 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4681 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4741 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4801 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4861 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4921 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4981 atgacccgca agcccggtgc ctgaggtacc tttaagacca atgacttaca aggcagctgt
     5041 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     5101 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     5161 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5221 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5281 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     5341 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     5401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5461 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     5521 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5581 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5641 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     5701 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5761 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5821 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5881 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5941 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6001 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6061 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6121 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6181 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6241 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6301 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     6361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7621 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7861 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaaga
     7921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     8281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     8341 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     8401 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     8461 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     8521 tcactaaagg gaacaaaagc tggagctgca agctt
//
