LOCUS       Exported                8371 bp ds-DNA     circular SYN 23-MAR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8371)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Dec 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8371
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    679..796
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    854..881
                     /label=bGH
     primer_bind     complement(871..893)
                     /label=Primer 4
     polyA_signal    882..1078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1079
                     /label=Barcode_Cloning_Site
     misc_feature    1145
                     /label=Linker
     misc_feature    1146..1165
                     /label=BFP_Perturb_Primer
     primer_bind     complement(1147..1178)
                     /label=Oligo_19
     misc_feature    1166
                     /label=GFP homology
     primer_bind     1175..1193
                     /label=Oligo_41
     primer_bind     1179..1193
                     /label=Oligo_42
     primer_bind     1179..1188
                     /label=Primer 1
     misc_feature    1180..1768
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1286..1297
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1695..1715
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1757..1801)
                     /label=Oligo_40
     primer_bind     complement(1757..1781)
                     /label=Primer 2
     primer_bind     1778..1825
                     /label=Primer 1
     CDS             complement(1805..2506)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2482..2527)
                     /label=Primer 2
     misc_feature    2537..2583
                     /label=chimeric intro
     intron          2584..3545
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     promoter        complement(3546..3821)
                     /label=chicken beta-actin promoter
     primer_bind     3587..3667
                     /label=Oligo_131
     primer_bind     3701..3710
                     /label=Oligo_131
     enhancer        3824..4188
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    4189..4201
                     /label=CMV_enhancer
     misc_feature    4292..4611
                     /label=U3
     LTR             4612..4792
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4854..5442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4934..4953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5613..6473)
                     /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"
     primer_bind     6236..6255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6474..6578)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6653..6672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6844..7223
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7225..7423
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             7441..7621
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7668..7793
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(8290..8371,1..152)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
       61 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
      121 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
      181 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
      241 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
      301 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
      361 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
      421 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
      481 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
      541 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
      601 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
      661 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      721 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      781 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
      841 gggcccgctc tagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
      901 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
      961 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     1021 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagct
     1081 cgaggccggt gtaatcagcc tagctacatg ccccatgacg atccacggat gtgctgtctg
     1141 aattcgcttg tgccccagtt tgctagggag gtccaggacg cggggaggcg gcccaaaggg
     1201 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     1261 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     1321 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     1381 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1441 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     1501 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1561 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1621 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1681 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1741 cacaaatttt gtaatccaga ggttgattgg atcgtacgtc ggcataagct ttgtagcggc
     1801 cctacttgta cagctcgtcc atgccgtaca ggaacaggtg gtggcggccc tcggagcgct
     1861 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca
     1921 cgtagtagta gccgggcagt tgcacgggct tcttggccat gtagatggtc ttgaactcca
     1981 ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc ttcagcacgc
     2041 cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta
     2101 cggggccgtc gggggggaag ttggtgccgc gcatcttcac cttgtagatc agcgtgccgt
     2161 cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag ttcatcacgc
     2221 gctcccactt gaagccctcg gggaaggaca gcttcttgta atcggggatg tcggcggggt
     2281 gcttcacgta cgccttggag ccgtacatga actgggggga caggatgtcc caggcgaagg
     2341 gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     2401 cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc atgcgcacct
     2461 tgaagcgcat gaactctttg atgacctcct cgcccttgct caccataccg gtagcgctag
     2521 ttacgactag caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc
     2581 tagagccgcc ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc
     2641 agccgtcccc ctgcggcagc cccgaggctg gagatggaga aggggacggc ggcgcggcga
     2701 cgcacgaagg ccctccccgc ccatttcctt cctgccggcg ccgcaccgct tcgcccgcgc
     2761 ccgctagagg gggtgcggcg gcgcctccca gatttcggct ccgccagatt tgggacaaag
     2821 gaagtccctg cgccctctcg cacgattacc ataaaaggca atggctgcgg ctcgccgcgc
     2881 ctcgacagcc gccggcgctc cggggccgcc gcgcccctcc cccgagccct ccccggcccg
     2941 aggcggcccc gccccgcccg gcacccccac ctgccgccac cccccgcccg gcacggcgag
     3001 ccccgcgcca cgccccgcac ggagccccgc acccgaagcc gggccgtgct cagcaactcg
     3061 gggagggggg tgcagggggg ggttacagcc cgaccgccgc gcccacaccc cctgctcacc
     3121 cccccacgca cacaccccgc acgcagcctt tgttcccctc gcagcccccc cgcaccgcgg
     3181 ggcaccgccc ccggccgcgc tcccctcgcg cacacgcgga gcgcacaaag ccccgcgccg
     3241 cgcccgcagc gctcacagcc gccgggcagc gcgggccgca cgcggcgctc cccacgcaca
     3301 cacacacgca cgcacccccc gagccgctcc cccccgcaca aagggccctc ccggagccct
     3361 ttaaggcttt cacgcagcca cagaaaagaa acgagccgtc attaaaccaa gcgctaatta
     3421 cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag
     3481 agccggggcg ggcggcgcga ggcggcgcgg agcggggcac ggggcgaagg caacgcagcg
     3541 actcccgccc gccgcgcgct tcgcttttta tagggccgcc gccgccgccg cctcgccata
     3601 aaaggaaact ttcggagcgc gccgctctga ttggctgccg ccgcacctct ccgcctcgcc
     3661 ccgccccgcc cctcgccccg ccccgccccg cctggcgcgc gccccccccc cccccgcccc
     3721 catcgctgca caaaataatt aaaaaataaa taaatacaaa attgggggtg gggagggggg
     3781 ggagatgggg agagtgaagc agaacgtggg gctcacctcg acccatggta atagcgatga
     3841 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     3901 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     3961 cttgatgtac tgccaagtgg gcagtttacc gtaaatagtc cacccattga cgtcaatgga
     4021 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     4081 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     4141 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     4201 gtcgagctga tctacgatgt agtaccttta agaccaatga cttacaaggc agctgtagat
     4261 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     4321 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     4381 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     4441 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     4501 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     4561 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     4621 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4681 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4741 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga
     4801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5281 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6061 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6121 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6181 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6241 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6301 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6361 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6421 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6481 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6541 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6601 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6661 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6721 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     6781 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     6841 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     6901 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6961 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     7021 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     7081 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     7141 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     7201 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     7261 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     7321 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     7381 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
     7441 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7501 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7561 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7621 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     7681 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     7741 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     7801 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     7861 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     7921 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     7981 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     8041 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     8101 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     8161 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     8221 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     8281 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     8341 agcgtcaatg acgctgacgg tacaggccag a
//
