LOCUS       sequence-13808-e        4850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4850)
  TITLE     2-series LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 4850)
  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: pET His6 StrepII TEV LIC cloning vector (2HR-T)
FEATURES             Location/Qualifiers
     source          1..4850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     122..139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    154..296
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(222..241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     382..404
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(398..589)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2047..2066
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2150..2169
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2150..2168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2257..2279
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2299..2316
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2332..2355
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2362..2382
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2507..2525)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2521..2568
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2934..2939,2940..2956,2957..2962))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2989..3007)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3075..3179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3180..3248,3249..4040)
                     /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(3398..3417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4211..4799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4700..4719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
       61 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      121 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
      181 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
      241 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
      301 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
      361 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
      421 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
      481 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
      541 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
      601 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
      661 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
      721 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
      781 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
      841 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
      901 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
      961 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     1021 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     1081 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     1141 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     1201 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     1261 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     1321 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     1381 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     1441 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     1501 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     1561 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     1621 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     1681 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     1741 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     1801 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc
     1861 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     1921 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     1981 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     2041 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     2101 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     2161 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa
     2221 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat
     2281 atacatatga aatcttctca ccatcaccat caccatggtt cttctgctag ctggagccac
     2341 ccgcagttcg aaaaaggcgc cgaaaacctg tacttccaat ccaatattgg aagtggataa
     2401 cggatccgcg atcgcggcgc gccacctggt ggccggccgg taccacgcgt gcgcgctgat
     2461 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2521 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     2581 actatatccg gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc
     2641 tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag
     2701 aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact
     2761 ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa
     2821 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     2881 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     2941 cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac
     3001 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3061 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3121 atccgctcat gagacaataa ccctgataaa tgcttcaata acattgaaaa aggaagagta
     3181 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3241 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3301 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3361 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3421 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3481 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3541 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3601 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3661 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3721 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3781 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3841 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3901 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3961 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4021 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4081 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4141 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4201 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4261 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4321 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4381 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4441 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4501 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4561 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4621 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4681 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4741 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4801 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
//
