LOCUS       sequence-421615-        6436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SUMO-GFP in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  AUTHORS   Peng Y, Huang Y, Kiessling F, Renn D, Rueping M
  TITLE     Nanobody-Based Lateral Flow Immunoassay for Rapid Antigen Detection 
            of SARS-CoV-2 and MERS-CoV Proteins.
  JOURNAL   ACS Synth Biol. 2025 Jan 9. doi: 10.1021/acssynbio.4c00592.
  PUBMED    39786915
REFERENCE   2  (bases 1 to 6436)
  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"; journalName: "ACS Synth 
            Biol. 2025 Jan 9. doi: 10.1021/acssynbio.4c00592."
COMMENT     Sequence Label: pET303_sumocodo_GFP_codoopti
FEATURES             Location/Qualifiers
     source          1..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             79..99
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             119..136
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             146..163
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1187..1204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1257..1275)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1271..1318
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1355..1810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1442..1461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1652..1673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1836..1940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1941..2009,2010..2801)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2159..2178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2972..3560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3461..3480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3714..3731
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3746..3885
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3811..3830)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3971..3993
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3987..4178)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4987..6069)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6031..6050
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6070..6147)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6353..6372
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag taataatttt gtttaacttt aagaaggagg tctagaatgg gtagcagcca
      121 tcaccatcat catcatagca gcggtctggt tccgcgtggt agccatatga gcgatagcga
      181 agttaatcaa gaagccaaac cggaagttaa gccggaagtg aaacctgaaa cacatattaa
      241 cctgaaagtg agtgatggca gcagcgaaat cttcttcaaa atcaaaaaaa caacaccgct
      301 gcgtcgtctg atggaagcat ttgcaaaacg tcagggtaaa gaaatggata gcctgcgttt
      361 tctgtatgat ggtattcgta ttcaggcaga tcagacaccg gaagatctgg atatggaaga
      421 taacgatatt atcgaagcac atcgtgagca gattggtgga tccatgagca aaggcgaaga
      481 actgtttacc ggtgttgttc cgattctggt tgaactggat ggtgatgtta atggccacaa
      541 attttcagtt agcggtgaag gcgaaggtga tgcaacctat ggtaaactga ccctgaaatt
      601 tatctgtacc accggcaaac tgccggttcc gtggccgaca ctggttacca cactgagcta
      661 tggtgttcag tgttttagcc gttatccgga tcatatgaaa cgccacgatt ttttcaaaag
      721 cgcaatgccg gaaggttatg ttcaagaacg taccatcttc ttcaaagatg acggcaacta
      781 taaaacccgt gccgaagtta aatttgaagg tgataccctg gtgaatcgca ttgaactgaa
      841 aggcatcgat tttaaagagg atggtaatat cctgggccac aaactggaat ataattataa
      901 tagccacaac gtgtacatca tggccgacaa acagaaaaat ggcatcaaag tgaatttcaa
      961 aacccgtcac aatatcgaag atggttcagt tcagctggca gatcattatc agcagaatac
     1021 cccgattggt gatggtccgg ttctgctgcc ggataatcat tatctgagca cccagagcgc
     1081 actgagcaaa gatccgaatg aaaaacgtga tcacatggtg ctgctggaat ttgttaccgc
     1141 agcaggtatt acccatggta tggatgaact gtataactaa ctcgagcacc accaccacca
     1201 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1261 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1321 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1381 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1441 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1501 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1561 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1621 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1681 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1741 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1801 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1921 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2041 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2101 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2161 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2221 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2341 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2461 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2521 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2581 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2641 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2701 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2761 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2941 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3541 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3601 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3661 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3721 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3781 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3841 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3901 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3961 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4021 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     4081 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     4141 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     4201 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     4261 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4321 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4381 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4441 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4501 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4561 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4621 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
     4681 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4741 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4801 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     4861 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     4921 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     4981 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5041 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5101 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5161 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5221 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5281 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5341 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5401 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5461 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5521 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5581 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5641 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5701 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5761 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     5821 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     5881 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     5941 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     6001 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     6061 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     6121 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     6181 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     6241 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6301 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6361 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6421 cggcgtagag gatcga
//
