LOCUS       sequence-208613-        3863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses lacZa-GFP fusion from constitutive proA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3863)
  AUTHORS   Davis JH, Rubin AJ, Sauer RT
  TITLE     Design, construction and characterization of a set of insulated 
            bacterial promoters.
  JOURNAL   Nucleic Acids Res. 2011 Feb;39(3):1131-41. doi: 10.1093/nar/gkq810. 
            Epub 2010 Sep 15.
  PUBMED    20843779
REFERENCE   2  (bases 1 to 3863)
  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"; doi: 
            "10.1093/nar/gkq810"; journalName: "Nucleic Acids Res"; date: 
            "2011-02"; volume: "39"; issue: "3"; pages: "1131-41"
COMMENT     Sequence Label: pSB3C5-proA-B0032-E0051
FEATURES             Location/Qualifiers
     source          1..3863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    33..54
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             226..399
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(246..263)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(246..262)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(255..277)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             436..453
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             466..1173
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     CDS             466..1173
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     primer_bind     complement(490..518)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1113..1133
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    1180..1200
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1236..1267
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      1778..2323
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2224..2243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2440..2457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3031..3690
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3097..3116)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3820..3849)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tacgtaatta gttagttagc ccttagtgac tcgaattcgc ggccgcttct agagcacagc
       61 taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg gataacttta
      121 cgggcatgca taaggctcgt aggctatatt cagggagacc acaacggttt ccctctacaa
      181 ataattttgt ttaactttta ctagagtcac acaggaaagt actagatgac catgattacg
      241 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      301 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      361 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgcggg cggcagcgaa
      421 ggcggcggca gcgaacatca tcatcatcat catggcagcg aacgtaaagg agaagaactt
      481 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      541 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
      601 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactaccct gacctatggt
      661 gttcaatgct ttgcgagata cccagatcat atgaaacagc atgacttttt caagagtgcc
      721 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
      781 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
      841 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
      901 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt
      961 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     1021 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     1081 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct
     1141 gggattacac atggcatgga tgaactatac aaataataat actagtagcg gccgctgcag
     1201 gagtcactaa gggttagtta gttagattag cagaaagtca aaagcctccg accggaggct
     1261 tttgactaaa acttcccttg gggttatcat tggggctcac tcaaaggcgg taatcagata
     1321 aaaaaaatcc ttagctttcg ctaaggatga tttctgctag agatggaata gactggatgg
     1381 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1441 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1501 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1561 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1621 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1681 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     1741 tccactgagc gtcagacccc ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     1801 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     1861 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     1921 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     1981 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     2041 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     2101 gagcgaactg cctacccgga actgagtgcc aggcgtggaa tgagacaaac gcggccataa
     2161 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     2221 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     2281 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     2341 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gcctcgttcg
     2401 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     2461 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     2521 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     2581 ccgctagcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat
     2641 cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt
     2701 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
     2761 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccacgttgt
     2821 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2881 actgtctgct tacataaaca gtaatacagg ggtgtttact agaggttgat cgggcacgta
     2941 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     3001 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcacggg atataccacc
     3061 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     3121 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     3181 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaacgctcac
     3241 ccggagtttc gtatggccat gaaagacggt gagctggtga tctgggatag tgttcaccct
     3301 tgttacaccg ttttccatga gcaaactgaa acgttttcgt ccctctggag tgaataccac
     3361 gacgatttcc ggcagtttct ccacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     3421 ctggcctatt tccctaaagg gtttattgag aatatgtttt ttgtctcagc caatccctgg
     3481 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     3541 ttcacgatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgatccag
     3601 gttcatcatg ccgtttgtga tggcttccat gtcggccgca tgcttaatga attacaacag
     3661 tactgtgatg agtggcaggg cggggcgtaa taatactagc tccggcaaaa aaacgggcaa
     3721 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg tttgccacct
     3781 gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc gaagaaaggc ccacccgtga
     3841 aggtgagcca gtgagttgat tgc
//
