LOCUS       40924_13895        7834 bp DNA     circular SYN 13-MAY-2021
DEFINITION  All-in-one doxycycline inducible lentiviral vector for expression of
            one or two genes using the P2A self-cleaving peptide..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7834)
  AUTHORS   Barger CJ, Branick C, Chee L, Karpf AR
  TITLE     Pan-Cancer Analyses Reveal Genomic Features of FOXM1 Overexpression 
            in Cancer.
  JOURNAL   Cancers (Basel). 2019 Feb 21;11(2). pii: cancers11020251. doi: 
            10.3390/cancers11020251.
  PUBMED    30795624
REFERENCE   2  (bases 1 to 7834)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7834)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancers
            (Basel)."; date: "2019-02-21"; pages: " 10.3390/cancers11020251"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     CDS             567..623
                     /codon_start=1
                     /label=P2A
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
                     /translation="ATNFSLLKQAGDVEENPGP"
     promoter        651..1161
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             1185..1877
                     /codon_start=1
                     /label=TurboRFP
                     /note="red fluorescent protein from Entacmaea quadricolor"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     CDS             1893..1946
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1947..2690
                     /codon_start=1
                     /label=rtTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(2733..2753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3151..3162)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     LTR             3367..3600
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3672..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3833..3968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3969..4039
                     /label=AmpR promoter
     CDS             4040..4897
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5071..5659
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5813..5830
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5947..5968
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5983..6013
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6021..6037
                     /label=lac operator
                     /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     6045..6061
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6082..6100
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6128..6354
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6355..6535
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6582..6707
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7200..7433
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7618..7662
                     /codon_start=1
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcgaattcgt cgacaccggt tccggagcca cgaacttctc tctgttaaag caagcaggag
      601 acgtggaaga aaaccccggt cctaggctgc agacgcgttg tacaggatcc ggggttgggg
      661 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
      721 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
      781 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
      841 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
      901 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
      961 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     1021 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     1081 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     1141 atcaccgacc tctctcccca gcaattcacc cgcgcgcggc caccatgagc gagctgatca
     1201 aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt
     1261 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaag atcaaggtgg
     1321 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
     1381 gcaaagcctt catcaaccac acccagggca tccccgactt ctttaagcag tccttccctg
     1441 agggcttcac atgggagaga atcaccacat acgaagacgg gggcgtgctg accgctaccc
     1501 aggacaccag cttccagaac ggctgcatca tctacaacgt caagatcaac ggggtgaact
     1561 tcccatccaa cggccctgtg atgcagaaga aaacacgcgg ctgggaggcc aacaccgaga
     1621 tgctgtaccc cgctgacggc ggcctgagag gccacagcca gatggccctg aagctcgtgg
     1681 gcgggggcta cctgcactgc tccttcaaga ccacatacag atccaagaaa cccgctaaga
     1741 acctcaagat gcccggcttc cacttcgtgg accacagact ggaaagaatc aaggaggccg
     1801 acaaagagac ctacgtcgag cagcacgaga tggctgtggc caagtactgc gacctcccta
     1861 gcaaactggg gcacagattc gaaggaagcg gagaaggtag aggttctctc ctcacttgtg
     1921 gtgatgttga agaaaaccct ggtccaatgt ctagactgga caagagcaaa gtcataaacg
     1981 gagctctgga attactcaat ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc
     2041 aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc
     2101 tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag
     2161 gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt gctctcctct
     2221 cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa
     2281 ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt
     2341 acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc
     2401 aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac
     2461 aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac
     2521 taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg accgacgccc
     2581 ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata
     2641 tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg taactaagta
     2701 aggatcgatc caagatatcg tattcttaac tatgttgctc cttttacgct atgtggatac
     2761 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2821 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2881 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2941 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3001 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3061 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3121 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3181 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3241 cggatctccc tttgggccgc ctccccgcct gtttcgcctc ggcgtccgga ctagaggtac
     3301 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     3361 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     3421 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     3481 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     3541 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     3601 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     3661 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3721 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3781 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     3841 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3901 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3961 tggaggcctt tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4021 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4081 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4141 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4201 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4261 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4321 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4381 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4441 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4501 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4561 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4621 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4681 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4741 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4801 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4861 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4921 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4981 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5041 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5101 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5161 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5221 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5281 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5341 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5401 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5461 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5521 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5581 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5641 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5701 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5761 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5821 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5881 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5941 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6001 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6061 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     6121 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     6181 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     6241 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     6301 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     6361 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6421 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6481 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     6541 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     6601 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     6661 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     6721 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     6781 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     6841 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     6901 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     6961 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7021 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     7081 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7141 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     7201 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     7261 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     7321 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     7381 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     7441 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     7501 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     7561 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     7621 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     7681 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     7741 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     7801 tcgtttcaga cccacctccc aaccccgagg ggac
//
