LOCUS       Exported                2725 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Level 1 cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    pUG-TP2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2725)
  AUTHORS   Kowarschik K, Hoehenwarter W, Marillonnet S, Trujillo M
  TITLE     UbiGate: a synthetic biology toolbox to analyse ubiquitination.
  JOURNAL   New Phytol. 2018 Mar;217(4):1749-1763. doi: 10.1111/nph.14900. Epub 
            2017 Nov 30.
  PUBMED    29194629
REFERENCE   2  (bases 1 to 2725)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2725
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     13..29
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13..29
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             25..348
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    42..98
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(99..116)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(99..115)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(108..130)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(324..343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(483..501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        569..673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             674..1534
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(892..911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1705..2293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2194..2213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2447..2464
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2640..2661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2676..2706
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2714..5
                     /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)."
     primer_bind     2719..16
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
       61 tcgactctag aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt
      121 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      181 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      241 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      301 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcataagg tctcagctta
      361 ctattgtctt ctaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      421 ttgtctgctc ccggcatccg cttacagaca agctgtgacc cacgaagtga gtggacgaaa
      481 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      541 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      601 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      661 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      721 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      781 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      841 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      901 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      961 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1021 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1081 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1141 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1201 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1261 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1321 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1381 tgagcgtggt tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1441 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1501 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1561 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1621 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1681 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1741 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1801 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1861 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1921 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1981 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2041 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2101 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2161 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2221 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2281 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2341 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2401 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2461 cgaggaagcg gatgagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2521 ttaatcactc tgtggttcag aagacatgca attaacttta agaaggagat ataccatgga
     2581 gaccgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2641 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2701 atgttgtgtg gaattgtgag cggat
//
