LOCUS       Exported                8518 bp ds-DNA     circular SYN 14-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8518)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 23, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8518
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1256
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1265..1306
                     /label=Primer 1
     CDS             1278..2072
                     /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"
     primer_bind     complement(2042..2081)
                     /label=Primer 2
     misc_feature    2088..2676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2559..2570)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2748..2981
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3059..3180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3220..3355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3376..3394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3404..3420)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3562..4017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4043..4147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4148..5008
                     /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"
     rep_origin      5179..5767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6055..6076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6091..6121
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6129..6145
                     /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     6153..6169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6190..6208
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6236..6462
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6463..6643
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6690..6815
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7308..7541
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        7948..8188
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    8198..8217
                     /label=CTRL g2
     misc_RNA        8218..8293
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    8349..8466
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgacgtac ggccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt
     1321 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     1381 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1441 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     1501 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     1561 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     1621 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     1681 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     1741 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     1801 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     1861 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     1921 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     1981 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2041 ccttctatcg ccttcttgac gagttcttct gaacgcgtta agtcgacaat caacctctgg
     2101 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2161 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2221 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2281 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     2341 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     2401 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2461 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     2521 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     2581 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     2641 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta
     2701 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     2761 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     2821 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     2881 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     2941 atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg
     3001 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     3061 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3121 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3181 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     3241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     3361 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     3421 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3481 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3541 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3601 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3661 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3721 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3781 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3841 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3901 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3961 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     4021 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4081 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4141 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4201 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4261 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4321 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4381 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4441 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4501 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4561 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4621 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4681 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4741 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4801 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4861 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4921 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4981 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5041 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5101 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5161 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5221 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5281 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5341 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5401 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5461 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5521 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5581 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5641 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5701 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5761 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5821 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5881 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5941 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6001 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6061 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6181 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
     6241 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
     6301 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
     6361 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
     6421 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
     6481 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6541 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6601 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     6661 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     6721 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     6781 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     6841 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     6901 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     6961 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     7021 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     7081 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     7141 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     7201 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     7261 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     7321 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     7381 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     7441 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     7501 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     7561 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     7621 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     7681 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     7741 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     7801 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     7861 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     7921 cacctcccaa ccccgagggg acccagagag ggcctatttc ccatgattcc ttcatatttg
     7981 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     8041 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     8101 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     8161 ttggctttat atatcttgtg gaaaggacga aacaccgcaa tcggcgacgt tttaaatgtt
     8221 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     8281 accgagtcgg tgctttttta agcttggcgt aactagatct tgagacaaat ggcagtattc
     8341 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     8401 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     8461 aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct cgaggggg
//
