LOCUS       sequence-325023-        3379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector that holds the linker for circuits with 2 TUs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3379)
  TITLE     Leonard Lab mMoClo Plasmids
REFERENCE   2  (bases 1 to 3379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pPD1152 pLink 2
FEATURES             Location/Qualifiers
     source          1..3379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      285..713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(372..391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     582..603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        736..932
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      783..918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     845..864
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1052..1173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1089..1108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1143..1162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1222..1238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1222..1238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1235..1257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1246..1262
                     /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)."
     promoter        complement(join(1270..1276,1277..1294,1295..1300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1315..1336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1453..1470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1624..2212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1704..1723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2383..3174,3175..3243))
                     /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     3006..3025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3244..3348)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattggaag acaaactaga
      181 ggatgcacat gtgaccgagg gattgtcttc gcatgctggg gatgcggtgg gctctatggc
      241 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
      301 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
      361 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
      421 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
      481 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
      541 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
      601 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
      661 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
      721 tggaatgtgt gtcagttagg ctcaattagt cagcaaccat agtcccgccc ctaactccgc
      781 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
      841 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
      901 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
      961 tcggaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     1021 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     1081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     1201 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2521 gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3361 aaaagtgcca cctgacgtc
//
