LOCUS       Exported                9332 bp ds-DNA     circular SYN 20-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9332)
  AUTHORS   Princeton Network
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 18, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9332
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
     LTR             836..1016
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1063..1188
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /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    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(2576..2593)
                     /label=OBA55
     misc_feature    2686..2999
                     /label=mU6
     protein_bind    2880..2913
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2917..2937
                     /label=GI library PCR 5' annealing
     misc_feature    3000..3019
                     /label=protospacer seq
     misc_feature    3020..3112
                     /label=constant region 1
     misc_feature    3142..3407
                     /label=hU6
     misc_feature    3408..3427
                     /label=protospacer seq
     misc_feature    3428..3522
                     /label=constant region 3
     misc_feature    complement(3540..3559)
                     
     promoter        3560..3783
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     complement(3619..3637)
                     /label=OBA421
     intron          3784..4722
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4500..4520)
                     /label=OBA57
     CDS             4743..5339
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             5349..5402
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(5376..5394)
                     /label=OBA59
     CDS             5415..6113
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     protein_bind    6133..6166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(6179..6198)
                     /label=OBA170
     misc_feature    6222..6810
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6693..6704)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6813..6829)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7086..7104)
                     /label=OBA46
     LTR             7339..7519
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7581..8169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7621..7638)
                     /label=OBA47
     CDS             complement(8340..9200)
                     /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(9201..9305)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
      841 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      901 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      961 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     1021 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     1081 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1141 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1201 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1261 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     1321 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1381 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1441 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1501 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc ccgctctaga ggacatgatg aggcaagaac
     2641 ctgaatgcag tctctcggag ggcgaagaat cctgcaggct cgagagatcc gacgcgccat
     2701 ctctaggccc gcgccggccc cctcgcacgg acttgtggga gaagctcggc tactcccctg
     2761 ccccggttaa tttgcatata atatttccta gtaactatag aggcttaatg tgcgataaaa
     2821 gacagataat ctgttctttt taatactagc tacattttac atgataggct tggatttcta
     2881 taacttcgta tagcatacat tatacgaagt tataaacagc acaaaaggaa actcacccta
     2941 actgtaaagt aattgtgtgt tttgagacta taagtatccc ttggagaacc accttgttgg
     3001 gacacccgag gcgcagcagg tttaagagct aagctggaaa cagcatagca agtttaaata
     3061 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttaattatac
     3121 ttaaggtgat gcaggctcga gccaaggtcg ggcaggaaga gggcctattt cccatgattc
     3181 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     3241 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     3301 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     3361 atttcgattt cttggcttta tatatcttgt ggaaagccac cttgttggag cgctcgggtg
     3421 ggactcagtt tcagagctaa gcacaagagt gcatagcaag ttgaaataag gctagtccgt
     3481 ttacaacttg aaaaagtggc acccgagtcg ggtgcttttt ttgcatgcgc ggcgcgccgt
     3541 ggataaccgt attaccgccg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     3601 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     3661 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     3721 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     3781 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
     3841 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
     3901 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
     3961 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     4021 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     4081 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     4141 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     4201 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     4261 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     4321 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     4381 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     4441 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     4501 aggcacctcg attagttctc gcgcttttgg agtacgtcgt ctttaggttg gggggagggg
     4561 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     4621 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     4681 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg agctagccca
     4741 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     4801 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     4861 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4921 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4981 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5041 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5101 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5161 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5221 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5281 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgccg
     5341 gatcgggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     5401 caccggtcgc caccagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
     5461 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     5521 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     5581 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
     5641 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     5701 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     5761 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
     5821 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
     5881 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac atcaagacca
     5941 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
     6001 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
     6061 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat taagaattcg
     6121 tcgagggacc taataacttc gtatagcata cattatacga agttatacat gtttaagggt
     6181 tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct ctggattaca
     6241 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     6301 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     6361 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     6421 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     6481 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6541 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6601 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     6661 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6721 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6781 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgagaccta
     6841 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     6901 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     6961 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     7021 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     7081 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg
     7141 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat
     7201 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     7261 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
     7321 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7381 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7441 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7501 gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7561 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7621 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7681 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7741 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7801 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7861 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7921 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7981 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     8041 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8101 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8161 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8221 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8281 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8341 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8401 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8461 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8521 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8581 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8641 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8701 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8761 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8821 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8881 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8941 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9001 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9061 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9121 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9181 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9241 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9301 ccgcgcacat ttccccgaaa agtgccacct ga
//
