LOCUS       pTP1172.        4672 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression plasmid of anti-mouse kappa chain nanobody 
            TP1170 (3x Cysteines).
ACCESSION   .
VERSION     .
KEYWORDS    pTP1172
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4672)
  AUTHORS   Pleiner T,  Bates M,  Gorlich D
  TITLE     A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies
  JOURNAL   J Cell Biol Dec 2017, jcb.201709115
REFERENCE   2  (bases 1 to 4672)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell Biol
            Dec 2017, jcb.201709115"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..45
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    53..69
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      908..1002
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1061..1873
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      2095..2181
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(2240..2261)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2277..3356)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3357..3434)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3518..3540)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3626..3766
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3781..3798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4631..4649
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatgag caagcatcac
      121 catcattcag gccatcacca taccggacac caccatcatt caggcagtca tcaccatacc
      181 ggcggatcat ccggaggtat gagctcagga tctaacggat ctacgggttc tggtactggt
      241 aatagcggga ccggaaccat gattaaagta aagactctga ccggcaagga aattgagatc
      301 gacatcgaac cgaccgacac catcgatcgt atcaaagaac gtgtggagga aaaagaaggc
      361 attcctccgg ttcagcagcg tctcatttac gcgggtaaac agcttgcaga tgacaaaacc
      421 gccaaagatt acaatatcga gggcggtagc gtactgcacc ttgttctggc cctgcgtggt
      481 ggcgccggta cctgcggatc ccaggtgcaa ttggtagagt gtgggggagg ctgggtgcag
      541 cctggggggt ctctgagact ctcctgtgca gcctctggat tcaccttcag tgacactgcc
      601 atgatgtggg tccgccaggc tccagggaag gggcgagagt gggtcgcagc tattgatact
      661 ggtggtggat acacatacta tgcagactcc gtgaagggcc gattcaccat ctccagagac
      721 aacgccaaga acacgctgta tctgcaaatg aacagcctga aaccggagga cacggcccga
      781 tactactgtg cgaagaccta ctccggtaat tactatagca actacacggt tgccaactat
      841 ggcaccacgg gccgggggac cctggtcaca gtctcatcat gttaagcttc ttaattagct
      901 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
      961 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gccatgaaaa
     1021 taaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1081 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1141 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1201 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1261 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1321 tatccgtact cctgatgatg cttggttact cacgactgcg atccccggca aaacagcatt
     1381 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1441 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1501 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1561 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcaca aacttttgcc
     1621 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1681 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1741 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1801 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1861 cgatgagttt ttctaagaat taattcatgg gcaaatatta tacgcaaggc gacaaggtgc
     1921 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     1981 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     2041 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
     2101 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2161 gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga taagctgtca
     2221 aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     2281 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2341 ggagaggcgg tttgcgtatt gggggccagg gtggtttttc ttttcaccag tgagacgggc
     2401 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2461 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     2521 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     2581 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     2641 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     2701 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     2761 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     2821 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     2881 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     2941 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3001 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3061 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3121 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     3181 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3241 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3301 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3361 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac
     3421 cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct
     3481 agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3541 catccgctta cagacaagct gtgaccgtct ccgggagcag acaagcccgt cagggcgcgt
     3601 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     3661 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     3721 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgacatt
     4621 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgaga aa
//
