LOCUS       sequence-456082-        4874 bp    DNA     circular SYN 15-APR-2025
DEFINITION  PZntA promoter driving expression of lacZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4874)
  AUTHORS   McNerney MP, Zhang Y, Steppe P, Silverman AD, Jewett MC, Styczynski 
            MP
  TITLE     Point-of-care biomarker quantification enabled by sample-specific 
            calibration.
  JOURNAL   Sci Adv. 2019 Sep 25;5(9):eaax4473. doi: 10.1126/sciadv.aax4473. 
            eCollection 2019 Sep.
  PUBMED    31579825
REFERENCE   2  (bases 1 to 4874)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2019-09-25"; volume: "5(9):eaax4473. doi"; pages: " 
            10.1126/sciadv.aax4473. eCollection 2019 Sep"
COMMENT     Sequence Label: pzntA_LacZ
FEATURES             Location/Qualifiers
     source          1..4874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             719..741
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             749..3823
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(769..786)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(769..785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(778..800)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3875..3893)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3889..3936
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3954..4769)
                     /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     4677..4696
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4761..4780
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa atctgtatct ctgataaaac ttgactctgg
      661 agtcgactcc agagtgtatc cttcggttaa tccacaacgg tttccctcta gaaataattt
      721 tgtttaactt taagaaggag atatacatat gaccatgatt acggattcac tggccgtcgt
      781 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      841 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      901 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
      961 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     1021 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     1081 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     1141 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgtta actcggcgtt
     1201 tcatctgtgg tgcaacgggc gctgggtcgg ttacggccag gacagtcgtt tgccgtctga
     1261 atttgacctg agcgcatttt tacgcgccgg agaaaaccgc ctcgcggtga tggtgctgcg
     1321 ctggagtgac ggcagttatc tggaagatca ggatatgtgg cggatgagcg gcattttccg
     1381 tgacgtctcg ttgctgcata aaccgactac acaaatcagc gatttccatg ttgccactcg
     1441 ctttaatgat gatttcagcc gcgctgtact ggaggctgaa gttcagatgt gcggcgagtt
     1501 gcgtgactac ctacgggtaa cagtttcttt atggcagggt gaaacgcagg tcgccagcgg
     1561 caccgcgcct ttcggcggtg aaattatcga tgagcgtggt ggttatgccg atcgcgtcac
     1621 actacgtctg aacgtcgaaa acccgaaact gtggagcgcc gaaatcccga atctctatcg
     1681 tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt gaagcagaag cctgcgatgt
     1741 cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg ctgaacggca agccgttgct
     1801 gattcgaggc gttaaccgtc acgagcatca tcctctgcat ggtcaggtca tggatgagca
     1861 gacgatggtg caggatatcc tgctgatgaa gcagaacaac tttaacgccg tgcgctgttc
     1921 gcattatccg aaccatccgc tgtggtacac gctgtgcgac cgctacggcc tgtatgtggt
     1981 ggatgaagcc aatattgaaa cccacggcat ggtgccaatg aatcgtctga ccgatgatcc
     2041 gcgctggcta ccggcgatga gcgaacgcgt aacgcgaatg gtgcagcgcg atcgtaatca
     2101 cccgagtgtg atcatctggt cgctggggaa tgaatcaggc cacggcgcta atcacgacgc
     2161 gctgtatcgc tggatcaaat ctgtcgatcc ttcccgcccg gtgcagtatg aaggcggcgg
     2221 agccgacacc acggccaccg atattatttg cccgatgtac gcgcgcgtgg atgaagacca
     2281 gcccttcccg gctgtgccga aatggtccat caaaaaatgg ctttcgctac ctggagagac
     2341 gcgcccgctg atcctttgcg aatacgccca cgcgatgggt aacagtcttg gcggtttcgc
     2401 taaatactgg caggcgtttc gtcagtatcc ccgtttacag ggcggcttcg tctgggactg
     2461 ggtggatcag tcgctgatta aatatgatga aaacggcaac ccgtggtcgg cttacggcgg
     2521 tgattttggc gatacgccga acgatcgcca gttctgtatg aacggtctgg tctttgccga
     2581 ccgcacgccg catccagcgc tgacggaagc aaaacaccag cagcagtttt tccagttccg
     2641 tttatccggg caaaccatcg aagtgaccag cgaatacctg ttccgtcata gcgataacga
     2701 gctcctgcac tggatggtgg cgctggatgg taagccgctg gcaagcggtg aagtgcctct
     2761 ggatgtcgct ccacaaggta aacagttgat tgaactgcct gaactaccgc agccggagag
     2821 cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg aacgcgaccg catggtcaga
     2881 agccggacac atcagcgcct ggcagcagtg gcgtctggct gaaaacctca gcgtgacact
     2941 ccccgccgcg tcccacgcca tcccgcatct gaccaccagc gaaatggatt tttgcatcga
     3001 gctgggtaat aagcgttggc aatttaaccg ccagtcaggc tttctttcac agatgtggat
     3061 tggcgataaa aaacaactgc tgacgccgct gcgcgatcag ttcacccgtg caccgctgga
     3121 taacgacatt ggcgtaagtg aagcgacccg cattgaccct aacgcctggg tcgaacgctg
     3181 gaaggcggcg ggccattacc aggccgaagc agcgttgttg cagtgcacgg cagatacact
     3241 tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag catcagggga aaaccttatt
     3301 tatcagccgg aaaacctacc ggattgatgg tagtggtcaa atggcgatta ccgttgatgt
     3361 tgaagtggcg agcgatacac cgcatccggc gcggattggc ctgaactgcc agctggcgca
     3421 ggtagcagag cgggtaaact ggctcggatt agggccgcaa gaaaactatc ccgaccgcct
     3481 tactgccgcc tgttttgacc gctgggatct gccattgtca gacatgtata ccccgtacgt
     3541 cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa ttgaattatg gcccacacca
     3601 gtggcgcggc gacttccagt tcaacatcag ccgctacagt caacagcaac tgatggaaac
     3661 cagccatcgc catctgctgc acgcggaaga aggcacatgg ctgaatatcg acggtttcca
     3721 tatggggatt ggtggcgacg actcctggag cccgtcagta tcggcggaat tccagctgag
     3781 cgccggtcgc taccattacc agttggtctg gtgtcaaaaa taagtcgacc ggctgctaac
     3841 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     3901 cttggggcct ctaaacgggt cttgaggggt tttttgctga aagccaattc tgattagaaa
     3961 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     4021 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     4081 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     4141 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     4201 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     4261 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     4321 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     4381 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     4441 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     4501 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     4561 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     4621 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     4681 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcttcgag
     4741 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     4801 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     4861 tgagacacaa cgtg
//
