LOCUS       Exported               12703 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    aMHC-Puro Rex-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12703)
  AUTHORS   Kita-Matsuo H, Barcova M, Prigozhina N, Salomonis N, Wei K, Jacot 
            JG, Nelson B, Spiering S, Haverslag R, Kim C, Talantova M, Bajpai R,
            Calzolari D, Terskikh A, McCulloch AD, Price JH, Conklin BR, Chen 
            HS, Mercola M
  TITLE     Lentiviral vectors and protocols for creation of stable hESC lines 
            for fluorescent tracking and drug resistance selection of 
            cardiomyocytes.
  JOURNAL   PLoS ONE. 2009  . 4(4):e5046.
  PUBMED    19352491
REFERENCE   2  (bases 1 to 12703)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE. 
            2009  . 4(4):e5046."
FEATURES             Location/Qualifiers
     source          1..12703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        50..353
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        354..557
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     508..528
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             571..751
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    798..923
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    974..1089
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     primer_bind     1165..1181
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1165..1181
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             6642..7241
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(6642..6661)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7138..7158
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             7985..8383
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8389..8977
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8442..8462)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8860..8871)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    9336..9369
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     LTR             9440..9620
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9670..9689)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(9903..9922)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10022..10044
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10082..10100)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10168..10272
                     /gene="bla"
                     /label=AmpR promoter
     CDS             10273..11133
                     /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"
     primer_bind     complement(10491..10510)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11304..11892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11793..11812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12046..12063
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(12133..12153)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12151..12448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        12249..12459
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      12318..12453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12380..12399
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    12569..12703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     12660..12679
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"