LOCUS       Exported                3041 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Start-Stop Assembly Level 234 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pStA234
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3041)
  AUTHORS   Taylor GM, Mordaka PM, Heap JT
  TITLE     Start-Stop Assembly: a functionally scarless DNA assembly system 
            optimized for metabolic engineering.
  JOURNAL   Nucleic Acids Res. 2018 Nov 20. pii: 5193345. doi: 
            10.1093/nar/gky1182.
  PUBMED    30462270
REFERENCE   2  (bases 1 to 3041)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3041
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      338..883
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     784..803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1000..1017
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1467..1486)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1478..2293
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1551..1570)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2336..2430
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    2480..2501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2516..2546
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2554..2570
                     /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     2559..2581
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2578..2594
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2578..2594
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2590..2913
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2607..2663
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2664..2681)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2664..2680)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2673..2695)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2889..2908)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      2943..3029
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcctcgtgat acgcaaatct ggagccggtg agcgcggtat cattgcagca ctggggccag
       61 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
      121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
      181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
      241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
      301 tccactgagc gtcagacccc ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
      361 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
      421 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
      481 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
      541 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
      601 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
      661 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
      721 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
      781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
      841 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
      901 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
      961 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     1021 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     1081 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     1141 ccgctagcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat
     1201 cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt
     1261 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
     1321 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccacgttgt
     1381 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1441 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1501 gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1561 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     1621 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1681 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1741 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
     1801 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1861 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1921 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     1981 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     2041 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2101 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2161 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2221 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2281 tgagtttttc taatcttggg ggaaattagc cgcagccgaa cgaccgagcg ccttggactc
     2341 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     2401 tgccgccggg cgttttttat tggtgagaat gaagacaatt acggagagag acccagtagg
     2461 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     2521 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     2581 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat
     2641 ccccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2701 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2761 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2821 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     2881 actctcagta caatctgctc tgatgccgca tagtttccgg tctcatactc gagaagtctt
     2941 ccatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     3001 tctttcgact gagcctttcg ttttatttga tgcccgaaag g
//
