LOCUS       sequence-207988-        4143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ecarin in Ecoli off a T7 promoter in the NEB control 
            cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4143)
  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 4143)
  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: pNL1 Ecarin
FEATURES             Location/Qualifiers
     source          1..4143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2121..2139)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2135..2182
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(2400..2988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2480..2499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3160..3951,3952..4020))
                     /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     3783..3802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4021..4125)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggagacc caagctggct
       61 agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc
      121 tgcagatatc caccatgata cagattcttc tggtaataat atgtttggcc gtattccctt
      181 atcaaggttg cagtattatt cttggtagtg gcaatgtcaa tgattacgaa gtggtgtacc
      241 cccaaaaggt tactgctctt ccaaaagggg ctgtgcagca gcccgagcag aaatacgagg
      301 acgcaatgca gtatgagttt gaggtgaaag gagagccagt agtgttacac ttggaaaaga
      361 ataaagaact gttctcggag gattactcag agacgcacta ctcttcggat gatagagaga
      421 tcaccacaaa tcctagcgta gaggaccatt gctattatca tggtcgcata caaaacgatg
      481 ctgagtcgac ggcctccatt tcggcgtgta atggactgaa aggtcatttt aaattgcggg
      541 gtgaaactta tttcatagag ccacttaaga ttcctgacag tgaggcgcat gccgtctaca
      601 agtacgagaa tatcgaaaat gaagacgagg cacctaaaat gtgtggagtt actcaggata
      661 attgggaaag tgatgagccg atcaaaaaaa cgcttggatt aatagttcct cctcacgaaa
      721 gaaagttcga gaagaagttt attgaattag tcgtcgtagt ggatcattca atggttacga
      781 aatacaacaa cgactcaacg gcgatccgta cctggatata tgaaatgctg aacacggtca
      841 atgagattta tttgcctttc aatataagag tagcattagt agggttagag ttttggtgta
      901 acggagactt aattaacgtc acttcgacgg cagacgatac attgcactca ttcggggagt
      961 ggcgggcgag tgatctgtta aaccgcaaaa gacacgatca tgctcagttg ttgacgaacg
     1021 tcaccttaga tcactctacg ctgggtatta cgtttgtgta tggaatgtgt aaatcggatc
     1081 ggtccgtgga gctgatttta gactattcaa acatcacgtt caacatggcc tacataattg
     1141 cacatgagat gggtcacagt ctgggaatgt tacatgacac gaaattctgc acttgtggag
     1201 ctaagccatg catcatgttt ggcaaagaaa gtatcccacc ccccaaagaa tttagttcat
     1261 gttcttatga tcagtataac aagtatttgt tgaagtataa tcctaaatgc atacttgacc
     1321 ctccgttgcg gaaagacata gcatcgccgg ctgtgtgtgg gaatgagatc tgggaggaag
     1381 gggaagaatg tgattgcggt tcaccagcgg actgtcgcaa cccctgttgt gacgctgcaa
     1441 catgcaagct gaaaccagga gcggagtgcg ggaatggtga gtgttgtgat aagtgcaaga
     1501 tacgcaaagc cggaactgag tgtagaccgg caagagacga ctgcgatgtg gctgaacact
     1561 gtactggaca atccgcggaa tgcccaagaa atgagtttca gcgcaacggt caaccatgtc
     1621 ttaataactc gggatactgc tacaatgggg actgtcctat tatgctgaac cagtgcattg
     1681 cgctgttctc gccctcagct acagttgcgc aggactcttg cttccaaaga aacttacaag
     1741 ggtcatacta cggctattgt accaaagaaa ttggatacta cggaaaaaga ttcccgtgtg
     1801 ctccgcagga tgtaaagtgc ggacgtctgt attgtctgga caattctttt aagaagaaca
     1861 tgagatgtaa gaacgattac tcctatgcgg acgagaacaa aggtatagtt gagccgggca
     1921 ccaagtgtga agacgggaag gtttgcatca atcgtaaatg tgtagatgta aacactgcct
     1981 actgaaggat cccgggaatt ctcgagtaag gttaacctgc aggaggcctt taattaaggt
     2041 ggtgcggccg cgctagcggt cccgggggat cgatccggct gctaacaaag cccgaaagga
     2101 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     2161 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggaagct tggcactggc
     2221 cgaccggggt cgagcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2281 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2341 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2401 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2461 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2521 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2581 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2641 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2701 tatcgtcttg agtccaaccc gctaagacac gacttatcgc cactggcagc agccactggt
     2761 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2821 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2881 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2941 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3001 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacagatccg ggattttggt
     3061 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3121 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3181 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3241 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3301 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3361 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3421 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3481 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3541 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3601 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3661 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3721 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3781 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3841 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3901 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3961 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4021 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4081 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4141 agt
//
