LOCUS       sequence-328984-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of proteins with a C-terminal thermostable GFP (TGP) tag 
            in insect cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  AUTHORS   Cai H, Yao H, Li T, Hutter CAJ, Li Y, Tang Y, Seeger MA, Li D
  TITLE     An improved fluorescent tag and its nanobodies for membrane protein 
            expression, stability assay, and purification.
  JOURNAL   Commun Biol. 2020 Dec 10;3(1):753. doi: 10.1038/s42003-020-01478-z.
  PUBMED    33303987
REFERENCE   2  (bases 1 to 5915)
  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: "Commun 
            Biol."; date: "2020-12-10"; volume: "3(1):753. doi"; pages: " 
            10.1038/s42003-020-01478-z"
COMMENT     Sequence Label: pFTSG
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(83..102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     292..313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        476..580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(581..649,650..1441)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(799..818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1612..2200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2101..2120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2354..2371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2505..2729
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2796..3329)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3499..3504,3505..3521,3522..3527))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4035..4083
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4108..4127
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4219..4328)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4352..4443
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4368..4385
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4532..4555
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5249..5272
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     polyA_signal    5399..5533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5436..5455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5490..5509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5562..5727
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      join(5911..5915,1..450)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
       61 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
      181 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
      241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct
      301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      421 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
      481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      541 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      601 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      661 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      721 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      781 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
      841 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      901 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      961 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1021 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1081 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     1141 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1201 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1261 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1321 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1381 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1441 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1501 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1561 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1621 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1681 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1741 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1801 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1861 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1921 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1981 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2041 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2101 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2161 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2221 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2281 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2341 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2401 tctccttacg catctgtgcg gtatttcaca ccgcatagac cagccgcgta acctggcaaa
     2461 atcggttacg gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg cggacaataa
     2521 agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa
     2581 tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat
     2641 ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg
     2701 tggccaaggg catggtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc
     2761 gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata
     2821 tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg
     2881 tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg
     2941 aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga
     3001 tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga
     3061 gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc
     3121 aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac
     3181 tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta
     3241 catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc
     3301 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     3361 tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc
     3421 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     3481 atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc
     3541 ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca
     3601 tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg
     3661 gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctt gcttcaggag
     3721 atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt
     3781 cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc tagctatatt
     3841 tctagtggtt ggctacgtac ccgtagtggc tatggcaggg cttgccgccc cgacgttggc
     3901 tgcgagccct gggccttcac ccgaacttgg gggttggggt ggggaaaagg aagaaacgcg
     3961 ggcgtattgg tcccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     4021 aaccccgcgt ttatgaacaa acgacccaac acccgtgcgt tttattctgt ctttttattg
     4081 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     4141 atctcccggt atcgcatgct atgcatcagc tgctagcacc atggctcgag atcccgggtg
     4201 atcaagtctt cgtcgagtga ttgtaaataa aatgtaattt acagtatagt attttaatta
     4261 atatacaaat gatttgataa taattcttat ttaactataa tatattgtgt tgggttgaat
     4321 taaaggtccg tatactccgg aatattaata gatcatggag ataattaaaa tgataaccat
     4381 ctcgcaaata aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata
     4441 aatattccgg attattcata ccgtcccacc atcgggcgca tctaaaatga gcggatccaa
     4501 gcttagcggc cgcgaattcc tcgagggtac cctggaagtg ctgtttcagg gcccgggcgg
     4561 tagtggaggt tccgcgagtg tcattaagcc ggagatgaaa attaagttgc gtatggaggg
     4621 ggccgtgaat gggcacaaat tcgttattga aggggagggg attggaaagc cgtatgaggg
     4681 tacgcaaacc ttggacttaa cagtagaaga aggggcacca ttgccttttt cttatgatat
     4741 tttgactccg gcgttccaat acgggaaccg tgctttcaca aagtatcccg aagatatccc
     4801 tgactatttc aagcaagcat ttcccgaggg ttacagttgg gagagaagta tgacctatga
     4861 ggatcaaggc atatgtattg ctacctctga cattactatg gagggagact gcttcttcta
     4921 tgagattcgt ttcgatggaa caaactttcc tcctaacggg ccagtgatgc agaaaaagac
     4981 gttgaagtgg gaaccgtcca cagaaaaaat gtatgtcgag gacggcgttc taaagggaga
     5041 cgtggaaatg gccttgctat tggaaggggg tggacactat cgttgtgatt tcaaaacgac
     5101 ctacaaggcc aaaaaagatg tgagactacc ggacgcccac gaggttgacc atcgtattga
     5161 aattttaagt cacgacaagg actataataa agtaagactg tacgaacacg cggaagcacg
     5221 ttacagcgga ggtggaagtg gtggtggcca ccatcaccat caccatcacc attaagtcga
     5281 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     5341 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     5401 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5461 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5521 tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata agtgaaatct
     5581 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     5641 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     5701 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat
     5761 caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac
     5821 agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     5881 cgcttatttg cagcctgaat ggcgaatggg acgcg
//
