LOCUS       sequence-468721-        5965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged Intracellular HABP4
            LCD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5965)
  TITLE     Mittag bacterial expression plasmids
REFERENCE   2  (bases 1 to 5965)
  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"
COMMENT     Sequence Label: pDest17_HABP4 LCD (1-413)
FEATURES             Location/Qualifiers
     source          1..5965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(152..176)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(229..247)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      243..290
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(656..661,662..678,679..684))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(711..729)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        797..901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(902..970,971..1762)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1120..1139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1933..2521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2422..2441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2675..2692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2707..2846
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2772..2791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2932..2954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2948..3139)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4601..4620
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4704..4723
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4704..4722
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4754..4776
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4796..4813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4823..4847
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4850..4870
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ccgcgcccgg gccgcggtgc aagaggtggt acacgcggcg gtcgtggccg cattcgtcgc
       61 gcagaaaatt atggtccgcg tgccgaagtg gttatgcagg atgtggcacc gaatccggat
      121 gatccggaag attttccggc actgagttaa aacccagctt tcttgtacaa agtggttgat
      181 tcgaggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      241 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      301 gaactatatc cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg
      361 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
      421 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
      481 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
      541 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
      601 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
      661 agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat
      721 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
      781 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      841 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      901 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      961 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1021 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1081 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1141 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1201 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1261 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1321 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1381 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1441 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1501 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1561 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1621 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1681 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1741 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1801 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1861 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1921 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1981 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2041 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2101 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2161 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2221 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2281 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2341 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2401 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2461 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2521 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2581 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2641 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2701 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     2761 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2821 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2881 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2941 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3001 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3061 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3121 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     3181 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     3241 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3301 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3361 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3421 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3481 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3541 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3601 cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg
     3661 acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt
     3721 gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca
     3781 ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag
     3841 ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc
     3901 cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg
     3961 aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat
     4021 cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc
     4081 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     4141 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     4201 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     4261 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     4321 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4381 ggctctcaag ggcatcggtc gatcgacgct ctcccttatg cgactcctgc attaggaagc
     4441 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     4501 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     4561 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     4621 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     4681 tcgagatctc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
     4741 ctctagaaat aattttgttt aactttaaga aggagatata catatgtcgt actaccatca
     4801 ccatcaccat cacctcgaat caacaagttt gtacaaaaaa gcaggctccg aaaacctgta
     4861 tttccagggc agtatgaaag gtgcactggg tagcccggtt gccgcagccg gtgcagctat
     4921 gcaggaaagt tttggttgtg tggttgcaaa tcgctttcat cagctgctgg atgatgaaag
     4981 tgatccgttt gatattctgc gcgaagcaga acgccgtcgt cagcagcagc tgcagcgtaa
     5041 acgtcgtgat gaagccgcag cagcagccgg tgcgggccct cgtggtggta gaagtccggc
     5101 aggtgcaagt ggccatcgtg ccggtgcagg tggtcgtcgt gaaagccaga aagaacgcaa
     5161 aagcctgccg gcaccggttg cccagcgtcc tgatagcccg ggtggtggtc tgcaggcccc
     5221 gggtcaaaaa cgtaccccgc gtcgtggtga acagcagggt tggaatgata gtcgcggccc
     5281 ggaaggtatg ctggaacgcg ccgaacgccg tagttatcgt gaatatcgtc cgtatgaaac
     5341 cgaacgccag gcagatttta ccgcagaaaa atttccggat gaaaaaccgg gtgaccgttt
     5401 tgatcgtgat cgcccgctgc gtggccgcgg tggtcctcgt ggtggaatgc gtggtcgcgg
     5461 tcgcggtggc cctggtaatc gtgtgtttga tgcctttgat cagcgtggta aacgtgaatt
     5521 tgaacgttat ggtggcaatg ataaaattgc agttcgtacc gaagataata tgggtggttg
     5581 cggtgtgcgc acctggggca gtggcaaaga taccagtgat gtggaaccga ccgccccgat
     5641 ggaagaaccg accgttgtgg aagaaagcca gggtaccccg gaagaagaaa gcccggcaaa
     5701 agttccggaa ctggaagttg aagaagaaac ccaggtgcag gaaatgaccc tggatgaatg
     5761 gaaaaatctg caggaacaga cccgtccgaa accggaattc aatattcgta aaccggaaag
     5821 taccgtgccg agtaaagccg tggttattca taaaagtaaa taccgcgatg atatggtgaa
     5881 agatgattat gaagatgata gccatgtttt tcgtaaaccg gccaatgata ttaccagcca
     5941 gctggaaatt aattttggta atctg
//
