LOCUS       pEPQD0KN0024_(pJL1-NET_ 2370 bp ds-DNA     circular     16-OCT-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     protein_bind    634..655
                     /label="CAP binding site"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     protein_bind    565..581
                     /label="lac operator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer_bind     454..471
                     /label="M13 Forward"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     complement(81..110)
                     /label="AN163\vector\amp\rev"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    472..528
                     /label="MCS"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer_bind     1652..1671
                     /label="pENTR-R"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     terminator      780..827
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    721..827
                     /label="3' UTR and T7 terminator from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    complement(153..689)
                     /label="lacZ expression casette"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    complement(552..675)
                     /label="Plac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             complement(845..1660)
                     /label="aminoglycoside O-phosphotransferase APH(3')-Ia, kanamycin resistance (kanR)"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     2271..2290
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     rep_origin      1782..2370
                     /label="pUC origin of replication (derivative of pBR322/pMB1/ColE1)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             complement(222..545)
                     /label="lacZ beta-galactosidase alpha peptide"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer_bind     455..471
                     /label="M13 fwd"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    108..122
                     /label="NET tag (N-terminal Expression Tag), MEKKI, from chloramphenicol acetyltransferase, minimizes secondary structure with RBS and 5' UTR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    787..801
                     /label="T7 term hairpin F"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     promoter        complement(589..619)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     complement(554..576)
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     promoter        28..45
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     RBS             78..100
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    join(761..2370,1..24)
                     /label="backbone from pJL1-sfGFP (Addgene #69496)"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     complement(541..557)
                     /label="M13 Reverse"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     227..246
                     /label="pRS-marker"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     1568..1587
                     /label="Kan-R"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    2307..2325
                     /label="QMD062 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer_bind     complement(541..557)
                     /label="M13 rev"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     complement(766..784)
                     /label="T7 Term"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    complement(808..822)
                     /label="T7 term hairpin R"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    93..99
                     /label="RBS"
                     /ApEinfo_revcolor=#800080
                     /ApEinfo_fwdcolor=#800080
     misc_feature    28..107
                     /label="T7 promoter and 5' UTR from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer_bind     440..462
                     /label="M13/pUC Forward"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(22..51)
                     /label="QMD051"
                     /note="sequence: tctGAAGACatgtgtccctatagtgagtcgtattaatttcg"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          46..67
                     /label="QMD052"
                     /note="sequence: tctGAAGACggacaccacaacggtttccctc"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(697..729)
                     /label="QMD532_pTRV2PEBV_partC_R"
                     /note="sequence: agaCCCGGGCACTtGAGACCtagctggcacgac"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          710..738
                     /label="QMD050"
                     /note="sequence: acaGAAGACatGGTCTCaGCTTcggctgctaacaaagccc"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          complement(81..106)
                     /label="QMD054"
                     /note="sequence: caGAAGACatTCTCacatttgtatatctccttcttaaagttaaac"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          104..154
                     /label="QMD663_lacZ_lifter_XbaI_AarI_F"
                     /note="sequence: gagGGTCTCtctagAAACgGcggcaggtggtcacagcttgtctgtaagcgg"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          complement(81..130)
                     /label="QMD053"
                     /note="sequence: caGAAGACatTCTCaCATTATTTTTTTCTCcatatgtatatctccttcttaaagttaaac"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(9..39)
                     /label="QMD416_WGpJL1_bbone_R"
                     /note="sequence: agaGAAGACttTTCGtcgcgggatcgcgttg"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          complement(923..946)
                     /label="QMD134_LETterm4_R, 61.2"
                     /note="sequence: ttttctccttcattacagaaacgg"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          116..145
                     /label="QMD044_ACCEPTOR_lacZ_F"
                     /note="sequence: agaGAAGACatgagaccgtcacagcttgtc"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          126..145
                     /label="QMD044 binding region (60.6)"
                     /note="sequence: tgagaccgtcacagcttgtc"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          complement(92..122)
                     /label="QMD379_24pJL1accept_to_CCAT_R"
                     /note="sequence: tctGAAGACtcATGGATATTTTTTTCTCcatatgtatatctccttct"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          126..145
                     /label="QMD380_25pJL1accept_to_CCAT_F"
                     /note="sequence: tctGAAGACtcACCATtGAGACCgtcacagcttgtc"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          28..47
                     /label="QMD060_T7_prom"
                     /note="sequence: taatacgactcactataggg"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          2304..2323
                     /label="QMD132_LETprom2_F, 61.4"
                     /note="sequence: gtttcgccacctctgacttg"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          750..780
                     /label="QMD417_WGpJL1_bbone_F"
                     /note="sequence: tgtGAAGACttTGCCaccgctgagcaataac"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          complement(9..24)
                     /label="QMD416 binding region 67.2 (16 bp)"
                     /note="sequence: tcgcgggatcgcgttg"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          2307..2325
                     /label="QMD062_pJL1_outside(BR322)_F"
                     /note="sequence: tcgccacctctgacttgag"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(690..738)
                     /label="QMD664_lacZ_lifter_XmaI_AarI_R"
                     /note="sequence: atgGGTCTCcccggCACTgagcgcaggtgtagctggcacgacaggtttc"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(765..783)
                     /label="QMD061_T7_term"
                     /note="sequence: ctagttattgctcagcggt"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          126..145
                     /label="QMD044"
                     /note="sequence: agaGAAGACatgagaccgtcacagcttgtc"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          2271..2294
                     /label="QMD131_LETprom1_F, 61.7, -3.89 self"
                     /note="sequence: gggaaacgcctggtatctttatag"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          126..145
                     /label="QMD378_24pJL1(26pEU)accept_to_CCAT_F"
                     /note="sequence: tctGAAGACtcCCATtGAGACCgtcacagcttgtc"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(892..919)
                     /label="QMD063_pJL1_outside(kanR)_R"
                     /note="sequence: tttcaaaaatatggtattgataatcctg"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(127..147)
                     /label="draft primer 61,6"
                     /note="sequence: cagacaagctgtgacGGTCTC"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          111..145
                     /label="QMD531_pTRV2PEBV_partC_F"
                     /note="sequence: agaGAAGACagAAACtGAGACCgtcacagcttgtc"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(723..740)
                     /label="QMD059"
                     /note="sequence: atGAAGACatCTCGAAGCtcgggctttgttagcagc"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          complement(830..856)
                     /label="QMD133_LETterm3_R, 61.4"
                     /note="sequence: gagtttttctaatcagaattggctttc"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(140..159)
                     /label="QMD129_MEKKIBsaI_lift_R, 61.3"
                     /note="sequence: ggcatccgcttacagacaag"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
ORIGIN
        1 aacgccagca acgcgatccc gcgaaattaa tacgactcac tatagggaca ccacaacggt
       61 ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg GAGAAAAAAA
      121 TAATGtGAGA CCgtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
      181 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
      241 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
      301 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
      361 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
      421 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag
      481 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
      541 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      601 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      661 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctaG GTCTCaGCTT
      721 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
      781 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaagccaatt
      841 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
      901 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
      961 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1021 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1081 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     1141 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1201 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     1261 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1321 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     1381 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     1441 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     1501 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     1561 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     1621 gcggcttcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     1681 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     1741 atcagagatt ttgagacaca acgtgagatc aaaggatctt cttgagatcc tttttttctg
     1801 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1861 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1921 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1981 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2041 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2101 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2161 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2221 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2281 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2341 tgctcgtcag gggggcggag cctatggaaa
//