LOCUS       pTIR_Antgra4_RFP        3638 bp ds-DNA     circular     11-APR-2025
DEFINITION  .
KEYWORDS    "Backbone" "Resistance" "Strain" "Gene of interest" "Glycerol Stock
            (date)"
COMMENT     Imported using the Genbank importer. File name:
            prfp-itr-9_piggybat.gb
FEATURES             Location/Qualifiers
     rep_origin      514..1102
                     /label="ori"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     1003..1022
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    complement(1127..1144)
                     /label="Rv primer"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1149..1280
                     /label="ITR5´"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(1158..1176)
                     /label="oITX15_H6_Antgra_4440_trim_ITR_Fw"
                     /note="sequence: GAATTCCGTCTCAGAGCTTTGTCCCATTGGGGCCGA"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          complement(1158..1178)
                     /label="oITX15_H7_Antgra_4440_trim_ITR_RV"
                     /note="sequence: GAATTCCGTCTCACATATTTGTCCCATTGGGGCCGA"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          1285..1303
                     /label="ITX12_H5.PB-512_bb_E3I.EcoRI._F_I"
                     /note="sequence: ATCGGAATTCCGTCTCATAACaaggatctgcgatcgctcc"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          1285..1303
                     /label="FWD_1/2GFPflip_F"
                     /note="sequence: ATCGCGTCTCAGAGCaaggatctgcgatcgctcc"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          1285..1304
                     /label="ITX1_B2_EF1p_Fw_Phos**"
                     /note="sequence: aaggatctgcgatcgctccg"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          complement(1285..1304)
                     /label="ITX1_G4_EF1_5’-ITR-junct-Rv"
                     /note="sequence: cggagcgatcgcagatcctt"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          complement(1285..1304)
                     /label="ITX15_A5_EF1_5' junction mapping REV_P5 miseq_FW"
                     /note="sequence: ACACTCTTTCCCTACACGACGCTCTTCCGATCTcggagcgatcgcagatcctt"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          1285..1305
                     /label="ROSA26_EF1a_Fw"
                     /note="sequence: CCATCTTCTAGAAAGACTGGAGTaaggatctgcgatcgctccgg"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          1285..1305
                     /label="EF1a_Fw"
                     /note="sequence: aaggatctgcgatcgctccgg"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          1285..1305
                     /label="TRACgRNA_EF1a_Fw"
                     /note="sequence: tacgaACCTCATGTCTAGCACAGTTTTGTaaggatctgcgatcgctccgg"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          1285..1305
                     /label="AAVS1gRNA_EF1a_Fw"
                     /note="sequence: tacgaCCAATCCTGTCCCTAGTGGCCCCaaggatctgcgatcgctccgg"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1285..1830
                     /label="EF1 promoter"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          1316..1353
                     /label="BspQ1-TET-on-Rv"
                     /note="sequence: ATCGaagcttgctcttccCATgggcagagcgcacatcgcccacagtccccgagaagtt"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1804..1861
                     /label="01.A6_P2A.RFP_E3I.AGGA_F"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="sequence: ATCGCGTCTCAaggagatgttgaagagaatcctggtcccatggccagctccgaggatg"
     misc_feature    complement(1812..1830)
                     /label="EF1 promoter rv"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    complement(1817..1857)
                     /label="14.G7_Ef1a.Esp3I.EcoRI_R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="sequence: tgcaggaattcgcagCGAGACGATATCGTAGGCGCCGGTCA"
     misc_feature    complement(1817..1874)
                     /label="14.G9_Ef1a.Esp3I.Esp3I.EcoRI_R"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="sequence: tgcaggaattcgcagCGAGACGtgctgaCGTCTCGgggcATATCGTAGGCGCCGGTCA"
     misc_feature    1829..1861
                     /label="01.A5_RFP_E3I.AAAT_F"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="sequence: TCGCGTCTCCAAATatggccagctccgaggatg"
     misc_feature    1843..2520
                     /label="mRFP"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     misc_feature    1851..1854
                     /label="GGov"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    1911..1930
                     /label="gRNA1"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    2609..2739
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(2646..2665)
                     /label="SV40pA-R"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          2697..2723
                     /label="oITX15_A3_3' RFP_P5_miseq_FW#2"
                     /note="sequence: ACACTCTTTCCCTACACGACGCTCTTCCGATCTGTTGTGGTTTGTCCAAACTCATCAATG"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2740..2767
                     /label="Orth-3p_GSP1"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          2740..2767
                     /label="ITX3_E1_Orth-3p_GSP1"
                     /note="sequence: GAATTGACTCAAATGATGTCAATTAGTC"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          2771..2792
                     /label="oITX1120_ITX14_I9_new_3' RFP_R_P5 miseq_FW"
                     /note="sequence: ACTGGAGTTCAGACGTGTGCTCTTCCGATCTCCAGAAGCTATCTGGTCTCCCTT"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(2771..2792)
                     /label="12.G2_SV40pA_TAAC_R"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="sequence: ATCGCGTCTCAGTTAAAGGGAGACCAGATAGCTTCTG"
     misc_feature    complement(2771..2792)
                     /label="seq SV40pA"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(2771..2792)
                     /label="ITX12_E1_seq SV40pA"
                     /note="sequence: AAGGGAGACCAGATAGCTTCTG"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    2791..2814
                     /label="GSP2?"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    2797..2925
                     /label="ITR3´"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    2849..2875
                     /label="Antgra_GSP2_80"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          2849..2875
                     /label="ITX3_E4_Antgra_GSP2_80"
                     /note="sequence: CAGACGTGTGCTCTTCCGATCTATGATCTCTTAAAATACCATAACTCGC"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer_bind     complement(2930..2948)
                     /label="pBRforEco"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(2967..2991)
                     /label="ITX16_B2_3ITR-ORTH-TTAA-Rv"
                     /note="sequence: tgacgtctaagaaaccattattatc"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     promoter        3016..3120
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             3121..343
                     /label="AmpR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer_bind     complement(3339..3358)
                     /label="Amp-R"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
ORIGIN
        1 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
       61 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      121 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      181 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      241 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      301 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
      361 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
      421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1141 ttgcTGCCTT AACCCTTTCG GCCCCAATGG GACAAATAAA TCCCAACATA CATTTCATAA
     1201 CATCTGCTGG GCGCTATCTG TCCCGTTTTT AATGGAAATT GTCATTAGCG GGTATTTTAA
     1261 GGTTTTAACA ATTTGGCAGC TATGaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     1321 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     1381 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     1441 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     1501 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     1561 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     1621 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     1681 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     1741 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     1801 ttacagatcc aagctgtgac cggcgcctac gatatcgcca ccatggccag ctccgaggat
     1861 gtcatcaaag agtttatgag atttaaggtc aagatggagg gaagcgtcaa cggacacgag
     1921 ttcgagattg agggagaagg agaaggccgg ccttacgagg gcacacaaac cgctaagctc
     1981 aaggtcacaa aaggaggacc cctccccttc tcctgggata ttctgagccc tcagttccag
     2041 tacggaagca aagcctatgt taaacaccct gccgacatcc ctgactatct gaagctctcc
     2101 ttccctgaag gcttcaagtg ggagagattc atgaacttcg aggacggagg cgtggtgaca
     2161 gtcacacaag atagcaccct ccaggacgga gagtttattt ataaggtgaa actcagagga
     2221 accaacttcc cctccgatgg ccctgtcatg caaaaaaaaa caatgggatg ggaagcctcc
     2281 accgagagaa tgtatcctga ggatggcgct ctgaaaggcg aaattaaaat gagactgaaa
     2341 ctcaaagacg gaggacacta cgatgccgag gtcaaaacaa cctacaaggc caagaaacaa
     2401 gtgcagctgc ctggcgccta catgactgat attaaactcg acattatcag ccataatggg
     2461 gactacacca tcgtggaaca atatgagaga gctgagggca gacatagcac aggcgctgga
     2521 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2581 tatgttgctc cttttacgct atgtgGTTAA CTTGTTTATT GCAGCTTATA ATGGTTACAA
     2641 ATAAAGCAAT AGCATCACAA ATTTCACAAA TAAAGCATTT TTTTCACTGC ATTCTAGTTG
     2701 TGGTTTGTCC AAACTCATCA ATGTATCTTA TCATGTCTGG AATTGACTCA AATGATGTCA
     2761 ATTAGTCTAT CAGAAGCTAT CTGGTCTCCC TTGAGCTCCC ACTCTCCCTG TATATATTTT
     2821 TTTTATTTTA ACTAGTATAT ATTGTCAAAT GATCTCTTAA AATACCATAA CTCGCAAAAA
     2881 CATCCTTAAA ATCATTAAAA ATAAAATTGG GTCCCAAAGG GTTAAAGGAg cctcgtgata
     2941 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3001 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3061 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3121 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3181 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3241 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3301 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3361 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3421 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3481 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3541 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3601 gatcgttggg aaccggagct gaatgaagcc ataccaaa
//