LOCUS       Exported                9570 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pLenti6/UbC/mSlc7a1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9570)
  AUTHORS   Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, 
            Yamanaka S
  TITLE     Induction of pluripotent stem cells from adult human fibroblasts by 
            defined factors.
  JOURNAL   Cell. 2007 Nov 30. 131(5):861-72.
  PUBMED    18035408
REFERENCE   2  (bases 1 to 9570)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2007 
            Nov 30. 131(5):861-72."
FEATURES             Location/Qualifiers
     source          1..9570
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        3..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."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        1803..3014
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2851..2870
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    3074..3098
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5003..5027)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5080..5121
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5158..5178)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5163..5492
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5343..5478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5540..5587
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5606..6004
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6091..6324
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6396..6517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6433..6452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6487..6506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6557..6692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6712..6731)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6713..6731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6741..6758)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6741..6757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6750..6772)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6899..7354
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6986..7005)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7196..7217
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7380..7484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7485..8345
                     /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(7703..7722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8516..9104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9005..9024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9258..9275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9392..9413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9428..9458
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9466..9482
                     /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     9471..9493
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9490..9506
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9490..9506
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9525..9545
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9527..9545
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"