LOCUS       pET-28a_PA1472_Cterm_Hi 5788 bp ds-DNA     circular     09-FEB-2023
DEFINITION  synthetic DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name:
            pET-28a_Chis_PA1472.txt NEBuilder-generated oligos (UPPERCASE =
            gene-specific, lowercase = overlap) PA1472_fwd:
            taagaaggagatataccatgATGAACTTTGTTTTTCCGC PA1472_fwd 3'Tm: 58.3
            3'Ta: 59.3 PA1472_rev: tggtggtggtggtgctcgagGCGATTACGCCAATGTTC
            PA1472_rev 3'Tm: 61.5  3'Ta: 59.3 pET-28a_Chis_fwd:
            CTCGAGCACCACCACCAC pET-28a_Chis_fwd 3'Tm: 68.9  3'Ta: 65.4
            pET-28a_Chis_rev: CATGGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAG
            pET-28a_Chis_rev 3'Tm: 64.4  3'Ta: 65.4
FEATURES             Location/Qualifiers
     primer_bind     2534..2553
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    2553..2575
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     protein_bind    2557..2573
                     /label="lac operator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(2581..2623)
                     /label="pET-28a_Chis_rev"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene-specific Tm: 64.4 Ta: 65.4"
                     /note="gene-specific primer: CATGGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAG"
     primer_bind     complement(2595..2623)
                     /label="pET28a Cterm Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     2604..2642
                     /label="PA1472_fwd"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="gene-specific Tm: 58.3 Ta: 59.3"
                     /note="overlap: taagaaggagatataccatg"
                     /note="gene-specific primer: ATGAACTTTGTTTTTCCGC"
     gene            2624..3175
                     /label="PA1472"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             2624..3202
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MNFVFPHPLDTPRLRLRTFREEDAAPLFAMMSDPEVMRYWNTPPWTTPAQAREAILRDSQALVDGEYLNLAIERREDGQLLGSCILFHFEKGSRRAELGYCLARAAQGRGYMGEALRRLLAFAFDEIDLNRLEAEIDPRNRPSAASLERLGFRQEGLLAQRWIVSGEVSDSALYGLLAEHWRNRLEHHHHHH*"
     primer_bind     complement(3158..3195)
                     /label="PA1472_rev"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="gene-specific Tm: 61.5 Ta: 59.3"
                     /note="overlap: tggtggtggtggtgctcgag"
                     /note="gene-specific primer: GCGATTACGCCAATGTTC"
     gene            3176..2623
                     /label="pET-28a_Chis"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     3176..3193
                     /label="pET-28a Cterm For"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3176..3193
                     /label="pET-28a_Chis_fwd"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="gene-specific Tm: 68.9 Ta: 65.4"
                     /note="gene-specific primer: CTCGAGCACCACCACCAC"
     primer_bind     3176..3196
                     /label="pET-28a For"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             3182..3199
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             3182..3199
                     /label="6x His Tag"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      3350..3805
                     /label="M13 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      3367..3673
                     /label="F1 ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(3901..4713)
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      4759..5441
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     5509..5529
                     /label="Seq Primer for Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     complement(5608..5628)
                     /label="2,900 R"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
ORIGIN
        1 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
       61 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
      121 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
      181 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
      241 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
      301 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
      361 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
      421 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      481 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
      541 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
      601 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
      661 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
      721 gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
      781 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
      841 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
      901 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
      961 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     1021 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     1081 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1141 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     1201 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     1261 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     1321 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     1381 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     1441 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     1501 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     1561 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     1621 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     1681 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     1741 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     1801 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     1861 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     1921 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     1981 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     2041 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     2101 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     2161 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     2221 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     2281 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     2341 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     2401 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     2461 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     2521 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     2581 ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgatgaact ttgtttttcc
     2641 gcatccgctg gatacaccgc gtctgcgtct gcgcaccttt cgtgaagaag atgcagcacc
     2701 gctgtttgca atgatgagcg atccggaagt tatgcgttat tggaataccc ctccgtggac
     2761 cacaccggca caggcacgtg aagcaattct gcgtgatagc caggcactgg ttgatggtga
     2821 atatctgaat ctggcaattg aacgtcgtga agatggtcag ctgctgggta gctgtattct
     2881 gtttcatttt gaaaaaggta gccgtcgtgc agaactgggt tattgtctgg cacgtgcagc
     2941 acaaggtcgt ggttatatgg gtgaagcact gcgtcgtctg ctggcatttg catttgatga
     3001 aattgatctg aatcgtctgg aagcagaaat cgatccgcgt aatcgtccga gcgcagcaag
     3061 cctggaacgt ctgggttttc gtcaagaagg cctgctggca cagcgttgga ttgttagcgg
     3121 tgaagttagc gatagcgcac tgtatggact gctggcagaa cattggcgta atcgcctcga
     3181 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3241 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3301 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     3361 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3421 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3481 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3541 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3601 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3661 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3721 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3781 aacaaaatat taacgtttac aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3841 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     3901 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     3961 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     4021 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     4081 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     4141 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     4201 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     4261 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     4321 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     4381 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     4441 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     4501 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     4561 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     4621 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     4681 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     4741 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4921 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4981 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5041 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5101 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5161 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5221 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5281 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5341 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5401 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5461 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5521 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5581 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     5641 gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata
     5701 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     5761 ccgccaacac ccgctgacgc gccctgac
//