LOCUS       Exported                9508 bp ds-DNA     circular SYN 20-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9508)
  AUTHORS   CUI Yan
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 6, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9508
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=5' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=HIV-1 RRE
                     /note="The Rev response element (RRE) of human 
                     immunodeficiency virus type 1 allows for Rev-dependent mRNA
                     export from the nucleus to the cytoplasm."
     misc_feature    2063..2180
                     /label=HIV-1 cPPT
                     /note="central polypurine tract from pol gene of human 
                     immunodeficiency virus type 1"
     CDS             complement(2245..2619)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        2713..2953
                     /label=U6 promoter
     primer_bind     2745..2768
                     /label=U6-seqF
     enhancer        3052..3355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3200..3219)
                     /label=CMV-seqR
     promoter        3356..3559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3651..4349
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     misc_feature    4375..4950
                     /label=IRES
     CDS             4951..5553
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5557..5649
                     /label=5' mir30 flank
     misc_feature    5653..5692
                     /label=Spacer 1
     misc_feature    5711..5750
                     /label=Spacer 2
     misc_feature    5778..6366
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element"
     misc_feature    6573..6806
                     /label=3'LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     rep_origin      complement(7235..7823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7994..8854)
                     /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(8855..8959)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8994..9186
                     /label=SV40 ori
     polyA_signal    9207..9341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     2761 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     2821 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     2881 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     2941 ttgtggaaag gacatcgatt ctttttttct agacgtatta ccgccatgca ttagttatta
     3001 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     3061 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     3121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     3181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     3241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     3301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     3361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     3421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     3481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     3541 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     3601 ctgttttgac ctccatagaa gacaccgact ctactagagg atctgccacc atggagagcg
     3661 acgagagcgg cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg
     3721 tggagttcga gctggtgggc ggcggagagg gcacccccga gcagggccgc atgaccaaca
     3781 agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg
     3841 gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg
     3901 ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc
     3961 tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtga
     4021 tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg
     4081 ccaccgtgga gcacctgcac cccatgggcg ataacgatct ggatggcagc ttcacccgca
     4141 ccttcagcct gcgcgacggc ggctactaca gctccgtggt ggacagccac atgcacttca
     4201 agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg
     4261 tggaggagga tcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga
     4321 ccccggatgc agatgccggt gaagaataat gtacaagtag cggccgcaaa ttccgcccct
     4381 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4441 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     4501 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     4561 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     4621 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     4681 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     4741 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     4801 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     4861 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     4921 tggttttcct ttgaaaaaca cgataatacc atggccaccg agtacaagcc cacggtgcgc
     4981 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     5041 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     5101 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5161 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5221 gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5281 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5341 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5401 gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5461 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5521 acctggtgca tgacccgcaa gcccggtgcc tgagtttgtt tgaatgaggc ttcagtactt
     5581 tacagaatcg ttgcctgcac atcttggaaa cacttgctgg gattacttct tcaggttaac
     5641 ccaacagaat tctcttctga agaaaatata tttcttttta ttcatagctc ttctcgaggt
     5701 aaccggatcc agttgtgttt taatgtatat taatgttact aatgtgtttt atttaaattg
     5761 agtcacgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5821 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5881 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5941 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6001 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6061 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6121 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     6181 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6241 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6301 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6361 ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag
     6421 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
     6481 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
     6541 tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag
     6601 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6661 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6721 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6781 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6841 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgtt taaacccgct
     6901 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     6961 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     7021 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     7081 agggggagga ttgggaagac aatagcaggg tatcagctca ctcaaaggcg gtaatacggt
     7141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7861 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7921 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7981 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8041 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8101 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8161 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8221 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8281 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8341 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8401 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8461 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8521 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8581 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8641 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8701 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8761 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8821 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8881 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8941 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gagttagtca
     9001 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     9061 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     9121 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     9181 aagcttcgtc gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat
     9241 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     9301 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta
     9361 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt
     9421 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
     9481 gttaaatatg tactacaaac ttagtagt
//
