LOCUS       sequence-424355-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  TITLE     2022-Q3 to 2023-Q1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6488)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 8DYS
FEATURES             Location/Qualifiers
     source          1..6488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..88
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  359..524
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    553..687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(577..596)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     631..650
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2580..2600)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2613..2630)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2689..2780)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(2747..2764)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(3132..3137,3138..3154,3155..3160)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3330..3863
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  3930..4154
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(4288..4305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4459..5047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4539..4558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5218..6009,6010..6078))
                     /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     5841..5860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6079..6183)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6209..6488,1..175))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6346..6367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
       61 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
      121 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
      181 tcgccattca ggctgcaaat aagcgttgat attcagtcaa ttacaaacat taataacgaa
      241 gagatgacag aaaaattttc attctgtgac agagaaaaag tagccgaaga tgacggtttg
      301 tcacatggag ttggcaggat gtttgattaa aaacataaca ggaagaaaaa tgccccgctg
      361 tgggcggaca aaatagttgg gaactgggag gggtggaaat ggagttttta aggattattt
      421 agggaagagt gacaaaatag atgggaactg ggtgtagcgt cgtaagctaa tacgaaaatt
      481 aaaaatgaca aaatagtttg gaactagatt tcacttatct ggttcggatc tcctaggcga
      541 tatcagtgat cagatccaga catgataaga tacattgatg agtttggaca aaccacaact
      601 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      661 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
      721 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
      781 gctgattatg atcctctagt acttctcgac aagcttcgtc atcaaatacc caattccaaa
      841 tcttccagct cctggagaag ggctgtttct ttctgaattt tctccaactg atttttttct
      901 agctgttttc cagttgctgc ttgttctttc agttgttcga ttgctttcag tttcttcttt
      961 aggttcttga tttttttgtc tatctcaggg tccccagaaa ttgactgaga gacagtgttt
     1021 cgtggtgtgc tctgtggggc aggagtaggt gccaaatctg gactcttgtc acttcttgct
     1081 tcctgctttg cagctttctt agcttcatgc ttcctttgat ttttaagagc tgtttttgat
     1141 aatggcttat cgtttcctga ttgtggtttc atattctgag gtggttcctc ttcatgcaat
     1201 ttggaattgg tgattggttt atttcttaaa gctgggggtc tataagctgt tgcaacttta
     1261 ggttcctcat tgggtacttc acttggaact gcttggtaag ttattgtttt tgctggaaat
     1321 attccatcca aaaatggctg ccaagaaacc tgccataatt ctgcatttga tggcacatca
     1381 tacttgtgca agatagagcc agtataatgc caaattttgt atccattatt aacccgtaac
     1441 ctgggagcac atgtagctgt taaaatatgc tcaccatccg ggcaccaagc aaaatatgta
     1501 gaatcagaag ccaccggttt agaaataagt ttgtagtttt tcacatccca cacttccatt
     1561 tgtcccctca gatttccaaa tccagctaat actaatatat gtccatgagg gctatagtag
     1621 gctgcattac gaggaccagt tccaaagtca aatacaggat cacatttcaa gttgaaaatt
     1681 gtcgctttgg caggcataaa accatataca gcacaaaact cagtagaact agaattccaa
     1741 actacatcat aaatggggcc attttttggt aattgcacta cagcactttc tccatttgtt
     1801 gcaatgtagt gtagagtttg ttctccatag taggaagctc ctgtcttgtc aacatctgtg
     1861 ctagctatta ccaacacagc agtagctttt ttattccaca gcattgtaac tttatctgcc
     1921 ttaaagaaac ttttattagc taaagctgca tgaggtccag caaagttggg gtactgatat
     1981 aatctaacaa atgaaggtgc acctttactt cctggaacat agacagccac cttgtatggt
     2041 tggggtccag gtgataatac aaagtcatta attttttgca aatgcaattt atttgcaatt
     2101 gtgttaaaat tgttgttttc aaagaagtga acttcattgt taacattgcg ggcacaaaga
     2161 gtttcatctt ctgaccagga tggacaccaa ttttgcattt ttttctggat gaaagatttc
     2221 aaacatgtcc cagttttcac atcataaagt tgtaggttgg gtatcccagc tgtgccatct
     2281 ttagaagtag tgtaaggctg ccacgttgcc aggacagtat ttttgggtga gaattcaagg
     2341 caaactgcct tcaggaggtc gaaggagtgc agtagtccct tgttagtgac actgataata
     2401 tttacttttt ctccattgcc ccaggcaaac aaggtcccat ccttactaaa gatacagact
     2461 ttgcaattct tcccagattc ccttggaaac actgtgcttt ctgtaaaatg tggtggtcca
     2521 ttcaccatgt acagtccttc tgatcctcgg actgtcaaga gcggcgtgga cggcgccatg
     2581 ccctggaaat acaagttttc tctgccgctg ctgtgatgat gatgatgatg catggttttc
     2641 ggaccgagat ccgcgcccga tggtgggacg gtatgaataa tccggaatat ttataggttt
     2701 ttttattaca aaactgttac gaaaacagta aaatacttat ttatttgcga gatggttatc
     2761 attttaatta tctccatgat ctattaatat tccggagtat acgtagccaa ccactagaac
     2821 tatagctaga gtcctgggcg aacaaacgat gctcgccttc cagaaaaccg aggatgcgaa
     2881 ccacttcatc cggggtcagc accaccggca agcgccgcga cggccgaggt cttccgatct
     2941 cctgaagcca gggcagatcc gtgcacagca ccttgccgta gaagaacagc aaggccgcca
     3001 atgcctgacg atgcgtggag accgaaacct tgcgctcgtt cgccagccag gacagaaatg
     3061 cctcgacttc gctgctgccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
     3121 cacgaaccca gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc
     3181 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     3241 ttttcatggc ttgttatgac tgtttttttg tacagtctat gcctcgggca tccaagcagc
     3301 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     3361 agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta
     3421 ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt
     3481 tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc gggaacttgc
     3541 tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt gttggcgctc
     3601 tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat atctatgatc
     3661 tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc aatctgctca
     3721 agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg
     3781 atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata
     3841 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc ccggccgcgg
     3901 agttgttcgg taaattgtca caacgccgcg aatatagtct ttaccatgcc cttggccacg
     3961 cccctcttta atacgacggg caatttgcac ttcagaaaat gaagagtttg ctttagccat
     4021 aacaaaagtc cagtatgctt tttcacagca taactggact gatttcagtt tacaactatt
     4081 ctgtctagtt taagacttta ttgtcatagt ttagatctat tttgttcagt ttaagacttt
     4141 attgtccgcc cacacccgct tacgcagggc atccatttat tactcaaccg taaccgattt
     4201 tgccaggtta cgcggctggt ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4261 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4321 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4381 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4441 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4501 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4561 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4621 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4681 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4801 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4861 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4921 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4981 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5041 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5101 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5161 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5221 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5281 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5341 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5401 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5461 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5521 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5581 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5641 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5701 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5761 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5821 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5881 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5941 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6001 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6061 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6181 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     6241 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6301 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6361 agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6421 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6481 cactaaat
//
