LOCUS       62056_20980        4411 bp DNA     circular SYN 24-MAY-2021
DEFINITION  Cloning vector pTarget-eGFP-neo, complete sequence.
ACCESSION   MW246027
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4411)
  AUTHORS   Dias MM, Vidigal J, Sequeira DP, Alves PM, Teixeira AP, Roldao A.
  TITLE     Insect High FiveTM cell line development using site-specific flipase
            recombination technology
  JOURNAL   G3 (Bethesda) (2021) In press
  PUBMED    33982066
REFERENCE   2  (bases 1 to 4411)
  AUTHORS   Dias MM, Vidigal J, Sequeira DP, Alves PM, Teixeira AP, Roldao A.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2020) Animal Cell Technology Unit, iBET, Instituto
            de Biologia Experimental e Tecnologica, Av. da Republica, Qta. do 
            Marques, ITQB/iBET building, Oeiras 2780-157, Portugal
REFERENCE   3  (bases 1 to 4411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda) (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-2020) Animal Cell Technology Unit, iBET, Instituto de 
            Biologia Experimental e Tecnologica, Av. da Republica, Qta. do 
            Marques, ITQB/iBET building, Oeiras 2780-157, Portugal"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..48
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        207..225
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             253..969
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             1075..1116
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             1126..1143
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    1161..1290
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     regulatory      complement(1434..1482)
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(1549..2340)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_recomb     2348..2395
     promoter        2444..2548
                     /label=AmpR promoter
     misc_feature    2733..3112
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     rep_origin      3581..4169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcgg atcctacctg
       61 gttctttccg cctcagaagc catacttaag ggctggctta actatgcggc atcagagcag
      121 attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac
      181 ataaccttat gtatcataca catacgattt aggtgacact atagaactcg agtgagcatc
      241 gatcccacga ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      301 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      361 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      421 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      481 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      541 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      601 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      661 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      721 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      781 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      841 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
      901 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      961 ctgtacaagt aaagcggccg caaagctagc ttcgatccag acatgataag atacattgat
     1021 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gcccgcggtt cgaaggtaag
     1081 cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac
     1141 cattgagttt atctgactaa atcttagttt gtattgtcat gttttaatac aatatgttat
     1201 gtttaaatat gtttttaata aattttataa aataatttca acttttattg taacaacatt
     1261 gtccatttac acactccttt caagcgcgtg ggatcggcga gtcagtgagc gaggaagcgg
     1321 agagcgccca atacgcaagg aaacagctat gaccatgtta atgcagctgg cacgacaggt
     1381 ttcccgactg gaaagcgggc agtgagcgga aggcccatga ggccagttaa ttaaacacaa
     1441 aaaaccaaca cacagatgta atgaaaataa agatatttta ttgcggccat cgtgatggca
     1501 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     1561 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     1621 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     1681 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     1741 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     1801 gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     1861 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     1921 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     1981 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     2041 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     2101 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg
     2161 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     2221 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     2281 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     2341 ccatggagaa gttcctattc cgaagttcct attcttcaaa aggtatagga acttcactag
     2401 tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2461 tttatttttt taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2521 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2581 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2641 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2701 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2761 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2821 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2881 cacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2941 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3001 caacatgggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3061 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3121 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3181 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3241 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3301 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3361 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3421 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3481 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3541 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3601 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3661 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3721 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3781 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3841 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3901 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3961 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4021 cggtcagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4081 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtggt
     4141 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4201 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4261 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga gcgaccgagc
     4321 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4381 cgcgttggcc gactcattaa tgcagggccg c
//
