LOCUS       Exported                9942 bp ds-DNA     circular SYN 23-JAN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 26, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9942
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    10..598
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(481..492)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             673..906
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        1111..1215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1216..2076
                     /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      2247..2835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3246..3267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3282..3312
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3320..3336
                     /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     3344..3360
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             3621..4254
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    4301..4426
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4923..5156
                     /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    5678..5795
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5844..7025
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6074..7016
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     misc_feature    7029..8540
                     /label=YAP1
     misc_feature    8560..9127
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    8577..9129
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             9144..9938
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 atagatccta atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
       61 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
      121 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
      181 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
      241 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
      301 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
      361 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
      421 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
      481 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
      541 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
      601 gagatccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
      661 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
      721 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
      781 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      841 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      901 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
      961 tatcagagag tgagaggccc gggttaatta aggaaagggc tagatcattc ttgaagacga
     1021 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1081 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1141 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1201 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1261 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1321 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1381 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1441 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1501 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1561 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1621 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1681 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1741 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1801 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1861 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1921 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1981 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2041 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2101 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2221 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2281 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2341 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2401 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2461 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2521 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2581 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2641 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2701 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2761 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2821 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2881 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2941 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3001 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3061 cattaatgca gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3121 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3181 gcaaaaagct ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3361 atgattacga atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3421 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
     3481 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
     3541 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
     3601 tgtactacaa acttagtagt tggaagggct aattcactcc caaagaagac aagatatcct
     3661 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     3721 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     3781 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     3841 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     3901 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     3961 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     4021 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     4081 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     4141 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     4201 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     4261 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     4321 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     4381 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     4441 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     4501 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     4561 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     4621 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     4681 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     4741 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     4801 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     4861 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     4921 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     4981 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     5041 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     5101 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     5161 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     5221 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     5281 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     5341 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     5401 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     5461 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     5521 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     5581 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     5641 tatcgccgaa ttcacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     5701 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     5761 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     5821 atccagtttg gactagtcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     5881 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     5941 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     6001 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     6061 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     6121 tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc
     6181 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     6241 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     6301 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     6361 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     6421 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     6481 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     6541 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     6601 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     6661 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     6721 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     6781 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     6841 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     6901 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     6961 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     7021 cgtgaagcat ggaccccggg cagcagccgc cgcctcaacc ggccccccag ggccaagggc
     7081 agccgccttc gcagcccccg caggggcagg gcccgccgtc cggacccggg caaccggcac
     7141 ccgcggcgac ccaggcggcg ccgcaggcac cccccgccgg gcatcagatc gtgcacgtcc
     7201 gcggggactc ggagaccgac ctggaggcgc tcttcaacgc cgtcatgaac cccaagacgg
     7261 ccaacgtgcc ccagaccgtg cccatgaggc tccggaagct gcccgactcc ttcttcaagc
     7321 cgccggagcc caaatcccac tcccgacagg ccagtactga tgcaggcact gcaggagccc
     7381 tgactccaca gcatgttcga gctcattcct ctccagcttc tctgcagttg ggagctgttt
     7441 ctcctgggac actgaccccc actggagtag tctctggccc agcagctaca cccacagctc
     7501 agcatcttcg acagtcttct tttgagatac ctgatgatgt acctctgcca gcaggttggg
     7561 agatggcaaa gacatcttct ggtcagagat acttcttaaa tcacatcgat cagacaacaa
     7621 catggcagga ccccaggaag gccatgctgt cccagatgaa cgtcacagcc cccaccagtc
     7681 caccagtgca gcagaatatg atgaactcgg cttcaggtcc tcttcctgat ggatgggaac
     7741 aagccatgac tcaggatgga gaaatttact atataaacca taagaacaag accacctctt
     7801 ggctagaccc aaggcttgac cctcgttttg ccatgaacca gagaatcagt cagagtgctc
     7861 cagtgaaaca gccaccaccc ctggctcccc agagcccaca gggaggcgtc atgggtggca
     7921 gcaactccaa ccagcagcaa cagatgcgac tgcagcaact gcagatggag aaggagaggc
     7981 tgcggctgaa acagcaagaa ctgcttcggc aggcaatgcg gaatatcaat cccagcacag
     8041 caaattctcc aaaatgtcag gagttagccc tgcgtagcca gttaccaaca ctggagcagg
     8101 atggtgggac tcaaaatcca gtgtcttctc ccgggatgtc tcaggaattg agaacaatga
     8161 cgaccaatag ctcagatcct ttccttaaca gtggcaccta tcactctcga gatgagagta
     8221 cagacagtgg actaagcatg agcagctaca gtgtccctcg aaccccagat gacttcctga
     8281 acagtgtgga tgagatggat acaggtgata ctatcaacca aagcaccctg ccctcacagc
     8341 agaaccgttt cccagactac cttgaagcca ttcctgggac aaatgtggac cttggaacac
     8401 tggaaggaga tggaatgaac atagaaggag aggagctgat gccaagtctg caggaagctt
     8461 tgagttctga catccttaat gacatggagt ctgttttggc tgccaccaag ctagataaag
     8521 aaagctttct tacatggtta ctagctcgag tctagaggat ccctcccccc cccctaacgt
     8581 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     8641 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     8701 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     8761 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     8821 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     8881 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     8941 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     9001 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     9061 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     9121 atgataatat ggccacacat atgatgattg aacaagatgg attgcacgca ggttctccgg
     9181 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     9241 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     9301 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     9361 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     9421 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     9481 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     9541 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     9601 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     9661 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     9721 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     9781 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     9841 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     9901 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat cg
//
