LOCUS       Exported                8868 bp ds-DNA     circular SYN 26-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Jun 26, 2021 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8868
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' 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=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2238..2617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2618..2821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    2846..3490
                     /label=sfGFP 1-10
     CDS             3518..3679
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             3707..3727
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        3787..4297
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     regulatory      4325..4334
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4337..4363
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4364..4375
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4376..4766
                     /label=MS2 coat binding protein (MCP)
     CDS             4775..4795
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4871..4936
                     /label=GFP11
     CDS             5009..5170
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             5198..5218
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5258..5846
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5729..5740)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5921..6154
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6359..6463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6464..7324
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7495..8083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8494..8515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8530..8560
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8568..8584
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8592..8608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagactcg aggcgttgac attgattatt gactagttat taatagtaat caattacggg
     2281 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2341 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2401 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2461 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2521 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2581 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2641 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2701 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2761 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2821 tctctggcta actaccggtg ccaccatgtc caagggagaa gagttgttta ctggcgtagt
     2881 ccctattctc gtggaactcg atggtgacgt gaatggccat aagttttctg tcagaggaga
     2941 gggagaaggc gatgccacca tcgggaaact cacgctgaaa ttcatctgta ccactggaaa
     3001 acttcccgtg ccttggccaa ccctcgtgac aacactcacc tacggggtgc aatgtttctc
     3061 tcggtacccg gaccatatga agaggcatga cttcttcaag agtgccatgc ccgagggtta
     3121 cgttcaggag cgcaccatct cttttaagga cgatggcaaa tataagacaa gagcagtcgt
     3181 caagttcgag ggtgatacac tcgttaaccg catcgagctc aaaggcaccg attttaagga
     3241 ggacggaaat atcctgggac acaaattgga gtacaacttc aacagtcaca acgtgtatat
     3301 tacagcagat aagcagaaga atggcataaa ggccaatttc acggtaagac ataatgtcga
     3361 ggatggcagt gtccagctgg cagaccacta ccagcaaaac acgcccattg gcgatggacc
     3421 tgttctcctg ccagacaacc actacctcag tacccaaaca gtcctgtcca aggaccctaa
     3481 tgagaaagga atcgatggtg gaggtcggac cgaagagtac aagcttatcc tgaacggtaa
     3541 aaccctgaaa ggtgaaacca ccaccgaagc tgttgacgct gctaccgcgg aaaaagtttt
     3601 caaacagtac gctaacgaca acggtgttga cggtgaatgg acctacgacg acgctaccaa
     3661 aaccttcacg gtaaccgaag gtggtggtag cggtggtggt actagtccca agaagaagcg
     3721 caaggtgtaa gcggccgccg ggtttattac agggacagca gagatccact ttggccgcgg
     3781 ctcgaggggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
     3841 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggactc gcacattctt
     3901 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
     3961 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
     4021 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
     4081 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
     4141 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
     4201 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
     4261 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccac
     4321 cggtgccacc atgggctacc cctacgacgt gcccgactac gccatcgaag gccgccatat
     4381 gctagccgtt aaaatggctt ctaactttac tcagttcgtt ctcgtcgaca atggcggaac
     4441 tggcgacgtg actgtcgccc caagcaactt cgctaacggg atcgctgaat ggatcagctc
     4501 taactcgcgt tcacaggctt acaaagtaac ctgtagcgtt cgtcagagct ctgcgcagaa
     4561 tcgcaaatac accatcaaag tcgaggtgcc taaaggcgcc tggcgttcgt acttaaatat
     4621 ggaactaacc attccaattt tcgccacgaa ttccgactgc gagcttattg ttaaggcaat
     4681 gcaaggtctc ctaaaagatg gaaacccgat tccctcagca atcgcagcaa actccggcat
     4741 ctacgcggat ccacctgttg ctactcctag catgcccaag aagaagcgca aggtgggttc
     4801 cggcgctggc ggaagccctg ggggcgggag cggtggctct ggttcttctg ctagtggcgg
     4861 ctcaacatct atggagaaga gagaccacat ggtgctgcta gagtacgtga ccgccgcagg
     4921 aatcacagac gcttcaggag gcggaggttc tggtggaggc ggtagtggcg gtggagggtc
     4981 aatcgatggt ggaggtcgga ccgaagagta caagcttatc ctgaacggta aaaccctgaa
     5041 aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt tcaaacagta
     5101 cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca aaaccttcac
     5161 ggtaaccgaa ggtggtggta gcggtggtgg tactagtccc aagaagaagc gcaaggtgta
     5221 agcggccgct ctggctctcg agtagggtaa tctagataat caacctctgg attacaaaat
     5281 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5341 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5401 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5461 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5521 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5581 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5641 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5701 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5761 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5821 gatctccctt tgggccgcct ccccgcctga gatcctttaa gaccaatgac ttacaaggca
     5881 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     5941 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6001 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6061 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6121 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     6181 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggcccgg gttaattaag
     6241 gaaagggcta gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6421 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     6721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8281 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt
     8341 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     8401 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc
     8461 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     8521 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     8581 acaatttcac acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt
     8641 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     8701 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
     8761 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
     8821 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt
//
