LOCUS       pET-28a_SpeG_Nterminal_ 5854 bp ds-DNA     circular     03-NOV-2021
DEFINITION  .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name: ChaC Nterminal
            His.txt synthetic DNA NEBuilder-generated oligos (UPPERCASE =
            gene-specific, lowercase = overlap) ChaC_fwd:
            tggtgccgcgcggcagccatATGCCAAGCGCCCACAGTG ChaC_fwd 3'Tm: 65.9  3'Ta:
            66.2 ChaC_rev: tggtggtggtggtgctcgagCTATTGTGCGGTCGGCTTCAG ChaC_rev
            3'Tm: 63.1  3'Ta: 66.2 pET-28a cut_fwd: CTCGAGCACCACCACCACCAC
            pET-28a cut_fwd 3'Tm: 67.3  3'Ta: 70.0 pET-28a cut_rev:
            ATGGCTGCCGCGCGGCAC pET-28a cut_rev 3'Tm: 71.6  3'Ta: 70.0
FEATURES             Location/Qualifiers
     primer_bind     2552..2571
                     /label="T7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    2571..2593
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     complement(2613..2641)
                     /label="pET28a Cterm Rev"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             2639..3259
                     /label="Translation 2639-3259"
                     /translation="MGSSHHHHHHSSGLVPRGSHMPSAHSVKLRPLEREDLRYVHQLDNNASVMRYWFEEPYEAFVELSDLYDKHIHDQSERRFVVECDGEKAGLVELVEINHVHRRAEFQIIISPEYQGKGLATRAAKLAMDYGFTVLNLYKLYLIVDKENEKAIHIYRKLGFSVEGELMHEFFINGQYRNAIRMCIFQHQYLAEHKTPGQTLLKPTAQ*"
     primer_bind     2679..2717
                     /label="gene-specific primer: ATGCCAAGCGCCCACAGTG"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(2681..2698)
                     /label="pET-28 Nterm His Rev"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     gene            2699..3259
                     /label="SpeG"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(3239..3279)
                     /label="gene-specific primer: CTATTGTGCGGTCGGCTTCAG"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     gene            3260..2698
                     /label="pET-28a cut"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     3260..3277
                     /label="pET-28a Cterm For"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3260..3280
                     /label="pET-28a For"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             3266..3283
                     /label="6x His Tag"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             3266..3283
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      3434..3889
                     /label="M13 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      3451..3757
                     /label="F1 ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(3985..4797)
                     /label="KanR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      4843..5525
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
ORIGIN
        1 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
       61 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
      121 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
      181 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
      241 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
      301 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
      361 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
      421 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
      481 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
      541 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
      601 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
      661 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
      721 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
      781 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
      841 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
      901 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
      961 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     1021 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     1081 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1141 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     1201 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     1261 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     1321 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     1381 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     1441 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     1501 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     1561 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     1621 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     1681 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     1741 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     1801 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     1861 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     1921 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     1981 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     2041 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     2101 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     2161 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     2221 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     2281 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     2341 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     2401 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2461 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2521 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     2581 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
     2641 gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat
     2701 gccaagcgcc cacagtgtta agctacgccc gctggagcgt gaagatttac gctatgtaca
     2761 tcaactcgac aataacgcca gtgtgatgcg ttactggttt gaggaaccct acgaagcctt
     2821 tgttgaactc tctgatctgt atgataagca tattcacgat cagagcgaac ggcgctttgt
     2881 ggtggaatgt gacggcgaaa aagccggtct ggtggagctg gtggaaatta accatgttca
     2941 tcgccgcgca gaatttcaga taattatctc cccggagtat caggggaaag gtctggcaac
     3001 ccgtgccgcc aaattagcaa tggactatgg ctttaccgtt ctcaatctct ataagctgta
     3061 tctgatcgtt gataaagaga atgaaaaagc gattcacatt taccgcaagc ttggcttttc
     3121 ggttgaaggt gaattgatgc acgagttctt tattaatggt caatatcgta atgccattcg
     3181 catgtgtata ttccagcatc agtatctggc agagcacaaa acaccgggtc agactctcct
     3241 gaagccgacc gcacaatagc tcgagcacca ccaccaccac cactgagatc cggctgctaa
     3301 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3361 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     3421 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3481 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3541 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3601 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3661 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3721 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3781 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3841 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact
     3901 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3961 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     4021 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     4081 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     4141 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     4201 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     4261 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     4321 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4381 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     4441 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     4501 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     4561 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     4621 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     4681 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     4741 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     4801 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     4861 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4921 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4981 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5041 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5101 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5161 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5221 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5281 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5341 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5401 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5461 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5521 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5581 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5641 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5701 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     5821 tgggtcatgg ctgcgccccg acacccgcca acac
//