LOCUS       Exported                6703 bp ds-DNA     circular SYN 15-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-HTNC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  AUTHORS   Peitz M, Pfannkuche K, Rajewsky K, Edenhofer F
  TITLE     Ability of the hydrophobic FGF and basic TAT peptides to promote 
            cellular uptake of recombinant Cre recombinase: a tool for efficient
            genetic engineering of mammalian genomes.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Apr 2. 99(7):4489-94.
  PUBMED    11904364
REFERENCE   2  (bases 1 to 6703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     388..410
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     472..491
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     533..552
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        533..551
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    552..576
                     /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)."
     promoter        592..701
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             722..739
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             830..1858
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             1865..1888
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1978..1997)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2043..2098
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2097..2116)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2186..2233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2246..3704
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2334..2706
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2363..3704)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      complement(3863..4450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3942..3961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4463..5323)
                     /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     5086..5105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     5389..6216
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5389..5513
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5551..6156)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        6278..6581
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctt cgggctgtaa
      241 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      301 gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct
      361 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
      421 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
      481 tggttattgt gctgtctcat cattttggca aagaattgga tcggaccgaa attaatacga
      541 ctcactatag gggaattgtg agcggataac aattccccgg agttaatccg ggacctttaa
      601 ttcaacccaa cacaatatat tatagttaaa taagaattat tatcaaatca tttgtatatt
      661 aattaaaata ctatactgta aattacattt tatttacaat caaaggagat ataccatggg
      721 ccatcaccat caccatcacg gcatgggcgc tgcaggtcgc aagaaacgtc gccaacgtcg
      781 ccgtccgcct gcaggcacta gtgtaagctt gaagaagaag aggaaggtgt ccaatttact
      841 gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa
      901 gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct
      961 tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc
     1021 cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt
     1081 aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc
     1141 acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt
     1201 tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt
     1261 tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg
     1321 gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat
     1381 ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag
     1441 caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat
     1501 ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa
     1561 tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt
     1621 tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc
     1681 ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc
     1741 aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca tgaactatat
     1801 ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattagct
     1861 cgagcaccac catcaccatc accatcacta agtgattaac ctcaggtgca ggctgcctat
     1921 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcgatct
     1981 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     2041 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2101 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     2161 tttggcaaca tatgcccata tgtaactagc ataacccctt ggggcctcta aacgggtctt
     2221 gaggggtttt ttgctgaaag catgcggagg aaattctcct tgaagtttcc ctggtgttca
     2281 aagtaaagga gtttgcacca gacgcacctc tgttcactgg tccggcgtat taaaacacga
     2341 tacattgtta ttagtacatt tattaagcgc tagattctgt gcgttgttga tttacagaca
     2401 attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg tatgttagag
     2461 cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa tcctcaatag
     2521 atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa ccgatggctg
     2581 gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt agcagcaatc
     2641 tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa
     2701 tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga ctcgacgtaa
     2761 acacgttaaa tagagcttgg acatatttaa catcgggcgt gttagcttta ttaggccgat
     2821 tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat tttgccatag
     2881 ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt gtagttgagc
     2941 tttttggaat tatttctgat tgcgggcgtt tttgggcggg tttcaatcta actgtgcccg
     3001 attttaattc agacaacacg ttagaaagcg atggtgcagg cggtggtaac atttcagacg
     3061 gcaaatctac taatggcggc ggtggtggag ctgatgataa atctaccatc ggtggaggcg
     3121 caggcggggc tggcggcgga ggcggaggcg gaggtggtgg cggtgatgca gacggcggtt
     3181 taggctcaaa tgtctcttta ggcaacacag tcggcacctc aactattgta ctggtttcgg
     3241 gcgccgtttt tggtttgacc ggtctgagac gagtgcgatt tttttcgttt ctaatagctt
     3301 ccaacaattg ttgtctgtcg tctaaaggtg cagcgggttg aggttccgtc ggcattggtg
     3361 gagcgggcgg caattcagac atcgatggtg gtggtggtgg tggaggcgct ggaatgttag
     3421 gcacgggaga aggtggtggc ggcggtgccg ccggtataat ttgttctggt ttagtttgtt
     3481 cgcgcacgat tgtgggcacc ggcgcaggcg ccgctggctg cacaacggaa ggtcgtctgc
     3541 ttcgaggcag cgcttggggt ggtggcaatt caatattata attggaatac aaatcgtaaa
     3601 aatctgctat aagcattgta atttcgctat cgtttaccgt gccgatattt aacaaccgct
     3661 caatgtaagc aattgtattg taaagagatt gtctcaagct cggaacgctg cgctcggtcg
     3721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3841 aaaaggccgc gttgctggcg tttttccata ggctccgccc cctgacgagc atcacaaaaa
     3901 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3961 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4021 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4081 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4201 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4261 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4321 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4381 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4441 aggatctcaa gaagatcctt tgttaccaat gcttaatcag tgaggcacct atctcagcga
     4501 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4561 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4621 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4681 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4741 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4801 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4861 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4921 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4981 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5041 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5101 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5161 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5221 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5281 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5341 attattgaag catttatcag ggttattgtc tcatgtccgc gcgtttcctg catcttttaa
     5401 tcaaatccca agatgtgtat aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc
     5461 tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat ttgtaattat tgaataataa
     5521 aacaattata aatgtcaaat ttgtttttta ttaacgatac aaaccaaacg caacaagaac
     5581 atttgtagta ttatctataa ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa
     5641 atcattttca aatgattcac agttaatttg cgacaatata attttatttt cacataaact
     5701 agacgccttg tcgtcttctt cttcgtattc cttctctttt tcatttttct cttcataaaa
     5761 attaacatag ttattatcgt atccatatat gtatctatcg tatagagtaa attttttgtt
     5821 gtcataaata tatatgtctt ttttaatggg gtgtatagta ccgctgcgca tagtttttct
     5881 gtaatttaca acagtgctat tttctggtag ttcttcggag tgtgttgctt taattattaa
     5941 atttatataa tcaatgaatt tgggatcgtc ggttttgtac aatatgttgc cggcatagta
     6001 cgcagcttct tctagttcaa ttacaccatt ttttagcagc accggattaa cataactttc
     6061 caaaatgttg tacgaaccgt taaacaaaaa cagttcacct cccttttcta tactattgtc
     6121 tgcgagcagt tgtttgttgt taaaaataac agccattgta atgagacgca caaactaata
     6181 tcacaaactg gaaatgtcta tcaatatata gttgctctag ttattaatag taatcaatta
     6241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6541 cttggcagta catctacgta ttagtcatcg ctattaccat gcatggtcga ggtgagcccc
     6601 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     6661 attttttaat tattttgtgc agcgatgggg gcgggggggg ggg
//
