LOCUS       sequence-207981-        5101 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 5101)
  TITLE     2-series LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 5101)
  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 Sumo TEV LIC cloning vector (2S-T)
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             108..130
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             150..167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             186..479
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             486..506
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(631..649)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      645..692
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1058..1063,1064..1080,1081..1086))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1113..1131)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1199..1303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1304..1372,1373..2164)
                     /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(1522..1541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2335..2923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2824..2843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3077..3094
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3109..3248
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3174..3193)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3334..3356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3350..3541)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4999..5018
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagtgccggc tccggagagc
       61 tctttaatta agcggccgcc ctgcaggact cgagttctag aaataatttt gtttaacttt
      121 aagaaggaga tatacatatg aaatcttctc accatcacca tcaccatggt tcttctatgg
      181 ctagcatgtc ggactcagaa gtcaatcaag aagctaagcc agaggtcaag ccagaagtca
      241 agcctgagac tcacatcaat ttaaaggtgt ccgatggatc ttcagagatc ttcttcaaga
      301 tcaaaaagac cactccttta agaaggctga tggaagcgtt cgctaaaaga cagggtaagg
      361 aaatggactc cttaagattc ttgtacgacg gtattagaat tcaagctgat cagacccctg
      421 aagatttgga catggaggat aacgatatta ttgaggctca cagagaacag attggtggga
      481 tcgaggaaaa cctgtacttc caatccaata ttggaagtgg ataacggatc cgcgatcgcg
      541 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
      601 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      661 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
      721 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
      781 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
      841 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
      901 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
      961 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     1021 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
     1081 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     1141 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1261 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
     1321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     1561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1681 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     1861 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2161 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2221 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2461 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3001 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3061 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3121 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     3181 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3241 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3301 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3361 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3421 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     3481 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     3541 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     3601 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     3661 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     3721 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     3781 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     3841 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     3901 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     3961 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     4021 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     4081 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     4141 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     4201 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     4261 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     4321 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     4381 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     4441 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     4501 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     4561 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     4621 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     4681 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     4741 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4801 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     4861 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     4921 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     4981 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     5041 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     5101 t
//
