LOCUS       sequence-208510-        3344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ATF1 in Ecoli off a T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3344)
  AUTHORS   Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, 
            Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
  TITLE     BioBits Explorer: A modular synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. 
            eCollection 2018 Aug.
  PUBMED    30083608
REFERENCE   2  (bases 1 to 3344)
  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: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 
            10.1126/sciadv.aat5105. eCollection 2018 Aug"
COMMENT     Sequence Label: pJL1 ATF1
FEATURES             Location/Qualifiers
     source          1..3344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     617..636
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        617..635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             667..689
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2329..2347)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2343..2390
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2408..3223)
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3131..3150
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3215..3234
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga
      661 aataattttg tttaacttta agaaggagat atacatatga atgaaattga tgaaaagaac
      721 caagccccag tacagcaaga atgcttaaaa gaaatgattc agaacggaca tgcccgtcgt
      781 atggggtcgg tcgaggacct gtatgtggcg cttaaccgtc agaaccttta tcgtaacttt
      841 tgtacatacg gggagctgag tgactactgc acacgtgacc aacttacact ggcgctgcgt
      901 gagatttgcc tgaaaaatcc tactctttta catatcgtgt tgcccacgcg ctggccgaac
      961 cacgaaaact actatcgcag ttccgagtat tactcgcgtc cacatcccgt ccacgactac
     1021 atctccgtat tacaagaatt gaaacttagt ggagtagtgc ttaatgaaca acctgagtac
     1081 tcagcagtga tgaagcagat cttagaagaa ttcaagaatt caaagggcag ttatacagcg
     1141 aaaattttta aacttacaac cacactgaca attccttact tcggcccgac tggacccagt
     1201 tggcgcctta tctgtcttcc ggaggaacat actgagaaat ggaagaaatt tattttcgtc
     1261 agcaaccatt gcatgtcgga cggacgcagt tcaattcatt ttttccatga tttacgcgac
     1321 gagttaaata atattaagac ccctcctaaa aagttagact atatttttaa gtacgaggag
     1381 gattaccagt tgcttcgtaa acttcccgaa ccaatcgaaa aggtgatcga tttccgccct
     1441 ccgtacttat ttatccccaa aagcctttta tcgggattca tttacaacca tcttcgtttc
     1501 agcagcaaag gggtctgtat gcgcatggac gacgtggaaa aaactgacga tgttgtaaca
     1561 gaaattatta acatttcccc gactgaattc caagccatta aggcgaacat taaatcaaat
     1621 atccaaggca aatgtacgat tacccccttt ctgcacgtgt gttggttcgt gagccttcac
     1681 aagtggggca aattttttaa gcccttaaat tttgagtggc ttacggatat tttcatcccc
     1741 gcagattgtc gttcccaatt acctgatgat gatgagatgc gtcagatgta ccgttacggt
     1801 gccaacgtag gtttcattga ctttacgcca tggatttcag aatccgacat gaacgataac
     1861 aaagaaaatt tctggcctct gattgaacat taccatgaag ttatttcaga agcactgcgt
     1921 aataagaaac accttcacgg attagggttc aacatccaag gatttgtaca aaagtacgtt
     1981 aacatcgaca aggtgatgtg cgatcgtgca attggcaaac gtcgtggcgg tacactgttg
     2041 tccaatgtgg gactgtttaa tcagctggag gagcctgacg ctaaatacag catttgtgac
     2101 cttgccttcg gtcagtttca aggcagctgg caccaagctt tttcattagg ggtgtgtagc
     2161 actaatgtaa aagggatgaa cattgtggtc gcgagcacga aaaatgttgt ggggtcacag
     2221 gaatcattag aagaactgtg cagtatttac aaagctttgt tgctgggtcc ataataagtc
     2281 gaccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     2341 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaagcca
     2401 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     2461 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     2521 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     2581 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     2641 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     2701 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     2761 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     2821 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     2881 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     2941 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     3001 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     3061 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     3121 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     3181 atcgcggctt cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     3241 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     3301 aacatcagag attttgagac acaacgtgag atcaaaggat cttc
//
