LOCUS       sequence-235967-        2179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly AB part for pTac (Adam Meyer's improved 
            pTac promoter selected for dynamic range and low cross-sensitivity; 
            unpublished, using strategy from https://doi.org/10.1101/285866)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2179)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2179)
  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: P61m
FEATURES             Location/Qualifiers
     source          1..2179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     85..103
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(107..150)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    153..174
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        join(210..215,216..231,232..238)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    244..263
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_feature    275..295
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(301..318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(472..1060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(552..571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1231..2022,2023..2091))
                     /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     1854..1873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(2092..2179,1..17))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
       61 attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa
      121 aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agagactagt
      181 gggtctcagg agaacgatcg ttggctgtgt tgacaattaa tcatcggctc gtataatgtg
      241 tggaattgtg agcgctcaca atttactaga gacctactag tagcggccgc tgcaggcttc
      301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag
      481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gcgacccacg
     1381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2161 atgtatttag aaaaataaa
//
