LOCUS       sequence-446699-        6621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ColA BBa_B0014 tetAp SD8 ScTrpRS rrnB1 proB SD8 Lum1PylRS 
            BBaB10014RNAItrpA+ J23100 SD8 MmPylRS T3Te-T7Te DHFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4421 to 4455)
  AUTHORS   Costello A, Peterson AA, Lanster DL, Li Z, Carver GD, Badran AH
  TITLE     Efficient genetic code expansion without host genome modifications.
  JOURNAL   Nat Biotechnol. 2024 Sep 11. doi: 10.1038/s41587-024-02385-y.
  PUBMED    39261591
REFERENCE   2  (bases 1 to 6621)
  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"; journalName: "Nat 
            Biotechnol. 2024 Sep 11. doi: 10.1038/s41587-024-02385-y."
COMMENT     Sequence Label: pAMC070n1a12
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..636)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(146..165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      659..686
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    931..947
                     /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        1712..1767
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1718..1736
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1748..1766
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      4226..4253
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4421..4455
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     terminator      5883..5912
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(5913..5940)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 aaacgtccta gaagatgcca ggaggatact tagcagagag acaataaggc cggagcgaag
       61 ccgtttttcc ataggctccg cccccctgac gaacatcacg aaatctgacg ctcaaatcag
      121 tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctga tggctccctc
      181 ttgcgctctc ctgttcccgt cctgcggcgt ccgtgttgtg gtggaggctt tacccaaatc
      241 accacgtccc gttccgtgta gacagttcgc tccaagctgg gctgtgtgca agaacccccc
      301 gttcagcccg actgctgcgc cttatccggt aactatcatc ttgagtccaa cccggaaaga
      361 cacgacaaaa cgccactggc agcagccatt ggtaactgag aattagtgga tttagatatc
      421 gagagtcttg aagtggtggc ctaacagagg ctacactgaa aggacagtat ttggtatctg
      481 cgctccacta aagccagtta ccaggttaag cagttcccca actgacttaa ccttcgatca
      541 aaccgcctcc ccaggcggtt ttttcgttta cagagcagga gattacgacg atcgtaaaag
      601 gatctcaaga agatcattta cggattcccg acaccaaaga ggacatccgg ttcacactgg
      661 ctcaccttcg ggtgggcctt tctgcgttta tatactagag agagaatata aaaagccaga
      721 ttattaatcc ggctttttta ttatttagac ttagggaccc ttgtgcttct caaatgcctg
      781 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tcagtgcacg
      841 gctaactaag cggcctgctg actttctcgc cgatcaaaag gcattttgct attaagggat
      901 tgacgagggc gtatctgcgc agtaagataa ttgtgagcgg ataacaatta gcagacaaga
      961 taattttgca cccagcaaac ttggtacgta aacgcatcgt ggcgaactgg tccgggacca
     1021 ccgagcattc ctaagctccg gttagccggg ttcaactccc gggttcgtcg ccaaaatttg
     1081 aaagtgctgt aaggcacaga ccacccaagg gggacggtcc ggcgaccagc aggcctctat
     1141 aagcctcgcc ttgcggggtt cgacaccccg gtctctcgcc agtttaaaag acatcggcgt
     1201 caagcggatg tctggctgaa aggcctgaag aatttggacc tgtgatcatg tagatcgaga
     1261 ggcttctaat ccatatcagc cgggttagat tcccggcggg tccgccatct cttacttgat
     1321 atggctttag tagcggtatc aatatcagca gtaaaataaa tttcccgatc ccgccctagc
     1381 tcagaggtag agcgtgcctc tctagaaggg catggtcccc ggttcaaatc ctgggggcgg
     1441 gaccaactac tttatgtagt ctccgccgtg tagcaagaaa ttgagaagtg aactggtggc
     1501 tcaatggtag agctgcacgc ttccgaacgt gttggttgca ggttcaagtc ctgtccagtt
     1561 caccaattta ttcaagacgc ttaccttgta agtgcaccca gtaggatccg tcgacctgca
     1621 gatccttagc gaaagctaag gatttttttt aagcttctgc cattcatccg agacttaggg
     1681 accctgcggc cgccattaat tcctaatttt tgttgacact ctatcattga tagagttatt
     1741 ttaccactcc ctatcagtga tagagaaaag tgaaatgaat agacctgcag gtgcagtaag
     1801 gaggaaaaaa aaatgtctaa cgatgaaacc gttgaaaaag ttacccagca ggtttctgaa
     1861 ttgaaatcta ccgatgttaa agaacaggtt gttaccccgt gggatgttga aggtggtgtt
     1921 gatgaacagg gtcgtgcgca gaacattgat tacgataaat tgattaaaca gttcggtacc
     1981 aaaccggtta acgaagaaac cttgaaacgt ttcaaacagg ttaccggtcg tgaaccgcat
     2041 catttcttgc gtaaaggttt gttcttctct gaacgtgatt tcaccaaaat tttggatttg
     2101 tacgaacagg gtaaaccgtt cttcttggtt accggtcgtg gtccgtcttc tgattctatg
     2161 catttgggtc atatgattcc gttcgttttc accaaatggt tgcaggaagt tttcgatgtt
     2221 ccgttggtta ttccgttgac cgatgatgaa aaattcttgt tcaaacataa attgaccatt
     2281 aacgatgtta aaaacttcgc gcgtgaaaac gcgaaagata ttattgcggt tggtttcgat
     2341 ccgaaaaaca ccttcatttt ctctgatttg cagtacatgg gtggtgcgtt ctacgaaacc
     2401 gttgttcgtg tttctcgtca gattaccggt tctaccgcga aagcggtttt cggtttcaac
     2461 gattctgatt gtattggtaa attccatttc gcgtctattc agattgcgtg tgcgttcccg
     2521 tcttctttcc cgaacgtttt gggtttgccg gataaaaccc cgtgtttgtg tccgtgtgcg
     2581 attgatcagg atccgtactt ccgtgtttgt cgtgatgttg cggataaatt gaaatactct
     2641 aaaccggcgt tgttgcattc tcgtttcttc ccggcgttgc agggttctac caccaaaatg
     2701 tctgcgtctg atgataccac cgcgattttc atgaccgata ccccgaaaca gattcagaaa
     2761 aaaattaaca aatacgcgtt ctctggtggt caggtttctg cggatttgca tcgtgaattg
     2821 ggtggtaacc cggatgttga tgttgcgtac cagtacttgt ctttcttcaa agatgatgat
     2881 gttttcttga aagaatgtta cgataaatac aaatctggtg aattgttgtc tggtgaaatg
     2941 aaaaaattgt gtattgaaac cttgcaggaa ttcgttaaag cgttccagga acgtcgtgcg
     3001 caggttgatg aagaaacctt ggataaattc atggttccgc ataaattggt ttggggtgaa
     3061 aaagaacgtt tggttgcgcc gaaaccgaaa accaaacagg aaaaaaaata aacttaatta
     3121 acggcactca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttgctgagg
     3181 agacttaggg accctcacag ctaacaccac gtcgtcccta tctgctgccc taggtctatg
     3241 agtggttgct ggataacttt acgggcatgc ataaggctcg taatatatat tcagggagac
     3301 cacaacggtt tccctctaca aataattttg tttaacttta cctgcaggtg cagtaaggag
     3361 gaaaaaaaaa tggatacccg tttgaccccg gcgcaggcgc agcgtattcg tgaaatgggt
     3421 ggtaccgttg atccgtcttt ggcgttctct tctgaagcgg aacgtgaatc tgcgttccag
     3481 cgtatttctg cggatttgca gggtgcgaac ttggcgaaaa ttcgtcgttg tgcggaagcg
     3541 ccggaacgtc atccgattgg ttctttggaa aacaccttgg cgtgtgcgtt ggcggcgaaa
     3601 ggtttcattg aagttaaaac cccgatgatg attccggcgg atggtttggt taaaatgggt
     3661 attgatgaat ctcatccgtt gtggaaccag gttttctggg ttggtccgaa aaaagcgttg
     3721 cgtccgatgt tggcgccgaa cattttcttc ttgggtcgtc atttgcgtcg ttctgttccg
     3781 gcgccgttgt tgttgttcga aattggtccg tgtttccgta aagaatctcg tggttctaac
     3841 catttggaag aattcaccat gttgaacttg ttcgaattgg cgccgcaggc ggatgcgacc
     3901 gaacgtttga aagaacatat tgcgaccgtt atgaacgcgg ttggtttgcc gtacgaattg
     3961 gttgttgaag gttctgaagt tttcggtacc accattgatg ttgaagttga tggtgttgaa
     4021 ttggcgtctg gtgcggttgg tccgttgccg atggataaac cgcatggtat taccgaaccg
     4081 tgggcgggtg ttggtttcgg tttggaacgt attgcgttga tgcgtaccaa agaacagaac
     4141 attaaaaaag ttggtcgttc tttggtttac gttaacggtg cgcgtattga tatttaaacg
     4201 gagccaatgt cctcagcgtc acactggctc aacttcgggt gggcctttct gcgtttatat
     4261 actagagaga gaatataaaa agccagatta ttaatccggc ttttttatta tttggatccg
     4321 atccggcaaa caaaccaccg ttggtagcgg tggttttttt gtttgaccgg taaaaaaaag
     4381 cccgctcatt aggcgggctg ggtatctgat tgctggtacc ttgacggcta gctcagtcct
     4441 aggtacagtg ctagcggtgc agtaaggagg aaaaaaaaat ggataaaaaa ccgttgaaca
     4501 ccttgatttc tgcgaccggt ttgtggatgt ctcgtaccgg taccattcat aaaattaaac
     4561 atcatgaagt ttctcgttct aaaatttaca ttgaaatggc gtgtggtgat catttggttg
     4621 ttaacaactc tcgttcttct cgtaccgcgc gtgcgttgcg tcatcataaa taccgtaaaa
     4681 cctgtaaacg ttgtcgtgtt tctgatgaag atttgaacaa attcttgacc aaagcgaacg
     4741 aagatcagac ctctgttaaa gttaaagttg tttctgcgcc gacccgtacc aaaaaagcga
     4801 tgccgaaatc tgttgcgcgt gcgccgaaac cgttggaaaa caccgaagcg gcgcaggcgc
     4861 agccgtctgg ttctaaattc tctccggcga ttccggtttc tacccaggaa tctgtttctg
     4921 ttccggcgtc tgtttctacc tctatttctt ctatttctac cggtgcgacc gcgtctgcgt
     4981 tggttaaagg taacaccaac ccgattacct ctatgtctgc gccggttcag gcgtctgcgc
     5041 cggcgttgac caaatctcag accgatcgtt tggaagtttt gttgaacccg aaagatgaaa
     5101 tttctttgaa ctctggtaaa ccgttccgtg aattggaatc tgaattgttg tctcgtcgta
     5161 aaaaagattt gcagcagatt tacgcggaag aacgtgaaaa ctacttgggt aaattggaac
     5221 gtgaaattac ccgtttcttc gttgatcgtg gtttcttgga aattaaatct ccgattttga
     5281 ttccgttgga atacattgaa cgtatgggta ttgataacga taccgaattg tctaaacaga
     5341 ttttccgtgt tgataaaaac ttctgtttgc gtccgatgtt ggcgccgaac ttgtacaact
     5401 acttgcgtaa attggatcgt gcgttgccgg atccgattaa aattttcgaa attggtccgt
     5461 gttaccgtaa agaatctgat ggtaaagaac atttggaaga attcaccatg ttgaacttct
     5521 gtcagatggg ttctggttgt acccgtgaaa acttggaatc tattattacc gatttcttga
     5581 accatttggg tattgatttc aaaattgttg gtgattcttg tatggtttac ggtgatacct
     5641 tggatgttat gcatggtgat ttggaattgt cttctgcggt tgttggtccg attccgttgg
     5701 atcgtgaatg gggtattgat aaaccgtgga ttggtgcggg tttcggtttg gaacgtttgt
     5761 tgaaagttaa acatgatttc aaaaacatta aacgtgcggc gcgttctgaa tcttactaca
     5821 acggtatttc taccaacttg taaacttaat taacggcact cctcagctgt aaccaactca
     5881 ctggctcacc ttcacgggtg agcctttctt cgcgcagaaa ggcccacccg aaggtgagcc
     5941 agtgtgatta catttatgcc catggcgttt aacctttctg ccaaatctgg taagagtagt
     6001 taatgttaga cgcgaaatcc tgggtgaaaa ccggacggaa gttagacgga atttccggga
     6061 agtaaacatc accttccggt tcaatatcaa tggtagaaat atgcaaggta tcaacctgat
     6121 caatcaaaga tttgtaaatt tcaccaccac cagaaacaat aacatgatcg gtaatttttt
     6181 tcaagttggt caacgcatct ttaatagacg ggaaaatcaa aacgttttcg ttatcagagg
     6241 tgaaagaaga acgggtaaca accgcgtatt tacggttcgg caacgcaccc atagaaccga
     6301 aggttttacg accaaccaac aaccactggt tgtaggtaat cgctttgaac aacaactgtt
     6361 cacctttcgc agaccacgga atatccggac cgttaccaat aacaccgttt ttagaaatcg
     6421 caaccatcaa agacaatttc ataattcttc ctcagaggtt aacattttat tagtaagcat
     6481 gctcgttttg tcacccgctg atgcttaccg ttaattaata taatcacttg tattaaatag
     6541 actttctcta aaaatacaag acactctgtt attacaaatc gtgcgtgccg tctatcctgc
     6601 ttgcaaggcc aaataggccg t
//
