LOCUS       pCA1.327_Level_1        2749 bp DNA     circular SYN 18-SEP-2023
DEFINITION  Level 1 Position 1 pUC19 acceptor vector.
ACCESSION   .
VERSION     .
KEYWORDS    pCA1.327
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2749)
  AUTHORS   McCormick Lab
  TITLE     Alistair McCormick Lab Plasmids 2023
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 2749)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2023 from SnapGene 7.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
FEATURES             Location/Qualifiers
     source          1..2749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..33)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(16..32)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(25..47)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(241..260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     391..413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(451..469)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        537..641
                     /gene="bla"
                     /label=AmpR promoter
     CDS             642..1502
                     /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(860..879)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1673..2261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2162..2181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2415..2432
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2581..2602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2617..2647
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2655..2671
                     /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     2660..2682
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2679..2695
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2679..2695
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(2691..2749,1..265)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCNLSTICS
                     DAA"
     misc_feature    join(2708..2749,1..15)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
       61 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      121 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
      181 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcaatctcag
      241 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
      301 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgacggtctc
      361 acgctgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
      421 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
      481 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      601 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      661 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      721 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      781 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      841 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      901 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      961 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1021 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1081 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1141 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1201 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1261 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1321 ggtttattgc tgataaatct ggagccggtg agcgtggttc tcgcggtatc attgcagcac
     1381 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1441 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1501 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1561 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2341 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2401 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga tgagcgccca atacgcaaac
     2461 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcggagtg
     2521 agaccgcagc tggcacgaca ggtttgccga ctggaaagcg ggcagtgagc gcaacgcaat
     2581 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2641 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2701 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggt
//
