LOCUS       40924_42534        6284 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTag 2.0, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6284)
  AUTHORS   Vidigal J, Fernandes B, Dias MM, Patrone M, Roldao A, Carrondo MJT.,
            Alves PM, Teixeira AP.
  TITLE     RMCE-based insect cell platform to produce membrane proteins 
            captured on HIV-1 Gag virus-like particles
  JOURNAL   Appl. Microbiol. Biotechnol. (2017) In press
  PUBMED    29143881
REFERENCE   2  (bases 1 to 6284)
  AUTHORS   Vidigal J.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2017) Animal Cell Technology Unit, Ibet, Av. da 
            Republica, ITQB building, Oeiras 2780-157, Portugal
REFERENCE   3  (bases 1 to 6284)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6284)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-OCT-2017) Animal Cell Technology Unit, Ibet, Av. da Republica, 
            ITQB building, Oeiras 2780-157, Portugal"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        443..990
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect
                     cell expression"
     protein_bind    1027..1074
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1101..1817
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1972..2027
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     polyA_signal    2442..2563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2591..3613)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(3704..3995)
                     /label=OpIE-1 promoter
                     /note="moderate constitutive baculovirus promoter for
                     insect cell expression"
     primer_bind     complement(4063..4079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4087..4103)
                     /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)."
     promoter        complement(4111..4141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4156..4177)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4465..5053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5227..6084)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6085..6189)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tccaatccaa ttcatgatga taaacaatgt atggtgctaa tgttgcttca
      481 acaacaattc tgttgaactg tgttttcatg tttgccaaca agcaccttta tactcggtgg
      541 cctccccacc accaactttt ttgcactgca aaaaaacacg cttttgcacg cgggcccata
      601 catagtacaa actctacgtt tcgtagacta ttttacataa atagtctaca ccgttgtata
      661 cgctccaaat acactaccac acattgaacc tttttgcagt gcaaaaaagt acgtgtcggc
      721 agtcacgtag gccggcctta tcgggtcgcg tcctgtcacg tacgaatcac attatcggac
      781 cggacgagtg ttgtcttatc gtgacaggac gccagcttcc tgtgttgcta accgcagccg
      841 gacgcaactc cttatcggaa caggacgcgc ctccatatca gccgcgcgtt atctcatgcg
      901 cgtgaccgga cacgaggcgc ccgtcccgct tatcgcgcct ataaatacag cccgcaacga
      961 tctggtaaac acagttgaac agcatctgtt cgaatttagg atccgactag ttatcgattt
     1021 accggtgaag ttcctattcc gaagttccta ttctctagaa agtataggaa cttcgaggag
     1081 ctcgatatca agcttctacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1141 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1201 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1261 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1321 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1381 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1441 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1501 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1561 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1621 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1681 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1741 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1801 tggacgagct gtacaagtaa ggcggccgca ctcctcaggt gcaggctgcc tatcagaagg
     1861 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     1921 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     1981 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     2041 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     2101 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     2161 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     2221 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     2281 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     2341 acgtgagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2401 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2461 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2521 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggccagcta
     2581 gcaggattca ttccttggct ctgggtcttg tggagggtct cctgtttcca gagtcagcca
     2641 gaacttcaac acatccatca gtccaaacag cagcagacct tcttgcaatt tgagttcttc
     2701 caacagttcc agcaccagac ctcacaatgg catcacatct tccttgtgcc caagcagcat
     2761 catcaaagtt tccatcaacc agagattgat acagttggtc caggccaatt ctgagcatgt
     2821 aggctctcag tctgggggaa ccagccagtt ctgggtgtct tctttcaaaa tatcttgttt
     2881 gttgttccat gcaagccagc caaggtctcc aaaaaaaaat gttggcaacc tcatattgag
     2941 aatctccaaa catggcttca gaccagtcaa tgactgcagt gattctgcca ttgtctgtca
     3001 gaacattgtt gcttccaaaa tcagcatgga ccaggtgtct gacttcagga caatcttctg
     3061 cccacagcat gagttcatcc agtgcctgag caacagaagc agaaactgtg tcatccatca
     3121 cagtctgcca gtgatagaca tgaggatcag caatggcaca aatgaaatcc ctccaagtgg
     3181 tgtactgacc aatgccttgg ggaccaaaag gaccaaatcc agaggtttgg ctcagatcag
     3241 ctgctgcaat ggcatccatt gcttcagcaa caggttgcag aacagctggc agctcagttt
     3301 cagggagatc ctggagagtg actccttgtg ctcttctgct gatgcagtag gtgagagatt
     3361 cagaaaattc tccaatgtcc agaacttctg gaattggcag agcagcagag gcaaagtgtc
     3421 tgtaaacata tctgtctttg taaaaaccat cagcacaaga attgaccctc agaacataac
     3481 ctcttcctcc aacatcaaaa gaaaaggctc tgctttcttc accttcagac agctgcatga
     3541 gatcagaaac agaatcaaat ttttcaatga gaaacttctc aacagaagtt gctgtcagtt
     3601 caggtttctt catctagcga attcgcaatt gattgaagtt cctattccga agttcctatt
     3661 cttcaaaagg tataggaact tctatgatca gctcgagctt aaggcaaagg tgctgcgcgc
     3721 gccgcagcca acttgcaact gaaacaaaaa ccaacatgaa cgtctattta taccaaccgg
     3781 atggcgaaca agataatgat gtaacctttt acatgcctca cgccaccaac ggcgttatcg
     3841 tttatttatt caaaatgggc gcggaagccg cgcccgatag gacgcggctg gtgagcggat
     3901 atgaggccgg ccgaccgatt ggcctgcggt tagcggtggg cgcgtgcaac gacgcgtttg
     3961 tgctaagctg cgtgcgcgcg ccgtgcgtgt ttcgcattgg attggatcgg atcccgggcc
     4021 cgtcgactgc agaggcctgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4081 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4141 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4201 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4261 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4321 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4381 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4441 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4501 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4561 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4621 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4681 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4741 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4801 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4861 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4921 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4981 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5041 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5101 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5161 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5221 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5281 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5341 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5401 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5461 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5521 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5581 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5641 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5701 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5761 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5821 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5881 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5941 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6001 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6061 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6121 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6181 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6241 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
