LOCUS       pLenti6.3-lifeAc        8539 bp DNA     circular SYN 22-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2024 from SnapGene Viewer 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        1..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2247..2450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     TATA_signal     2436..2442
     protein_bind    2452..2470
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2473..2491
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2532..2556
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2557..2592
                     /label=MCS 1
                     /note="multiple cloning site, part 1"
     misc_feature    2593
                     /label=overhang+Kpn1
     misc_feature    2594
                     /label=T changed to G for Kozacs seq restauration
     misc_feature    2600..2650
                     /label=LifeAct
     misc_feature    2672..3382
                     /label=mNeonGreen
     misc_feature    3383..3385
                     /label=2nd stop codon
                     /note="Inserted by me. "
     misc_feature    3386
                     /label=XhoI+overhang
     misc_feature    3387..3396
                     /label=MCS 2
                     /note="multiple cloning site, part 2"
     protein_bind    complement(3400..3424)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3477..3518
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3542..4130
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element"
     promoter        4145..4461
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4312..4447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4509..4556
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4575..4973
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5060..5293
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5365..5486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5868..6323
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6349..6453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6454..6522,6523..7314)
                     /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      7485..8073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcttacat aacttacggt aaatggcccg cctggctgac
     1981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     2041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     2281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     2401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
     2461 gtgatagaga tctccctatc agtgatagag atcgtcgact agtccagtgt ggtggaattc
     2521 tgcagatatc aacaagtttg tacaaaaaag caggctggcg ccggaaccaa ttcagtcgac
     2581 tggatccggt accgccacca tgggcgtggc cgacctgatc aagaagttcg agagcatcag
     2641 caaggaggag ggggatccac cggtcgccac catggtgagc aagggcgagg aggataacat
     2701 ggcctctctc ccagcgacac atgagttaca catctttggc tccatcaacg gtgtggactt
     2761 tgacatggtg ggtcagggca ccggcaatcc aaatgatggt tatgaggagt taaacctgaa
     2821 gtccaccaag ggtgacctcc agttctcccc ctggattctg gtccctcata tcgggtatgg
     2881 cttccatcag tacctgccct accctgacgg gatgtcgcct ttccaggccg ccatggtaga
     2941 tggctccggc taccaagtcc atcgcacaat gcagtttgaa gatggtgcct cccttactgt
     3001 taactaccgc tacacctacg agggaagcca catcaaagga gaggcccagg tgaaggggac
     3061 tggtttccct gctgacggtc ctgtgatgac caactcgctg accgctgcgg actggtgcag
     3121 gtcgaagaag acttacccca acgacaaaac catcatcagt acctttaagt ggagttacac
     3181 cactggaaat ggcaagcgct accggagcac tgcgcggacc acctacacct ttgccaagcc
     3241 aatggcggct aactatctga agaaccagcc gatgtacgtg ttccgtaaga cggagctcaa
     3301 gcactccaag accgagctca acttcaagga gtggcaaaag gcctttaccg atgtgatggg
     3361 catggacgag ctgtacaagt aatagctcga gatatctaga cccagctttc ttgtacaaag
     3421 tggttgatat ccagcacagt ggcggccgct cgagtctaga ggggccgcgg ttcgaaggta
     3481 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggttag taatgatcga
     3541 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3601 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3661 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3721 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3781 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3841 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3901 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3961 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4021 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4081 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggcgatgg
     4141 taccggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4201 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4261 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4321 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4381 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4441 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4501 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4561 tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     4621 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     4681 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     4741 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     4801 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     4861 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     4921 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacaat
     4981 tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     5041 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     5101 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     5161 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5221 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     5281 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     5341 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     5401 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5461 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     5521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5581 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5641 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     5701 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5761 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5821 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     5881 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5941 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6001 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6061 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     6121 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     6181 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     6241 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     6301 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     6361 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6421 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6481 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6541 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6601 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6661 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6721 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6781 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6841 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6901 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6961 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7021 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7081 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7141 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7201 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7261 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7321 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7381 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7621 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8101 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8161 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     8221 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     8281 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     8341 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8401 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8461 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     8521 aagctggagc tgcaagctt
//
