LOCUS       sequence-383917-        6945 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 6945)
  TITLE     SGC Toronto 2022-Q1 to Q2
REFERENCE   2  (bases 1 to 6945)
  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: PATL2_G288-Y543_BV_bio
FEATURES             Location/Qualifiers
     source          1..6945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(176..181,182..198,199..204))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    712..760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     785..804
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(831..1796)
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             complement(834..1796)
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     promoter        complement(1833..1942)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1966..2057
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1982..1999
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2108..2152
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2954..2971
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3106..3240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3143..3162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3197..3216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3269..3434
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3618..4072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3914..3935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4098..4202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4203..4271,4272..5063)
                     /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(4421..4440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5234..5822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5723..5742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5976..5993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6127..6351
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
        1 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
       61 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
      121 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
      181 acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg
      241 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga
      301 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg
      361 caaggtgctg tgcacggatc tgcccttgct tcaggagatc ggtagacctc ggccgtcgcg
      421 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg
      481 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacgtacccg
      541 tagtggctat ggcagggctt gccgccccga cgttggctgc gagccctggg ccttcacccg
      601 aacttggggg ttggggtggg gaaaaggaag aaacgcgggc gtattggtcc caatggggtc
      661 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
      721 acccaacacc cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
      781 ggtattgtct ccttccgtgt ttcagttagc ctcccccatc tcccggtact tcatttttct
      841 gcactacgca gggatatttc accgcccatc cagggtttta ttattccatc ctgctcaagt
      901 aataaagccc cctgtttgtc tattccgcgt gaaatgccaa atatttcttt atcaccaatg
      961 ataagtttca ctgggcgatt aataaaatta tccagctttt cccagcgcga cagataaggt
     1021 gccaatcctt cttgttcgaa gagttccaac gcagcacgta attcacgtat tagcatggcc
     1081 gccaacgtat tacgatcgag attgatcccc gcttcctgca gcgtgatcca cccctgatta
     1141 acgacactct cttcaacacg gcgcattgcc atgttgatcc cggctccaat gactatttgc
     1201 gccgcatcgc cagttttgcc agtcagctcc accagaatgc ctgccagctt gcgatcctgc
     1261 agatagaggt cattaggcca tttaacacga actttatctg cacccagctt gcgtaatact
     1321 tccgccatca cgataccgat aaccagactt aaaccaatcg ccgccgccgg gccttgttcc
     1381 agacgccaga acatcgacaa atataagttt gcgccaaaag gcgaaaacca tttccgaccc
     1441 cggcgaccac ggccagcctg ctggtattct gcaatgcaag catcgcccga tttaagctct
     1501 ccgatacgat caagaaggta ctgattcgtg gagtcaatca ctggcagcac ggctacacta
     1561 ccgccatcca gctgacccaa tatctgttta gcattaagta actggatagg ctcaggcagg
     1621 ctgtatcctt tacccggaac ggtaaagaca tcaacgcccc agtcacgcag tgtctgaatg
     1681 tgtttattaa tagccgcccg gctcattccc agcgtttcac ccaactgctc gccagagtga
     1741 aattcaccgt tcgctaacag ggcaatcaat ttcagtggca cggtgttatc cttcatggct
     1801 cgagatcccg ggtgatcaag tcttcgtcga gtgattgtaa ataaaatgta atttacagta
     1861 tagtatttta attaatatac aaatgatttg ataataattc ttatttaact ataatatatt
     1921 gtgttgggtt gaattaaagg tccgtatact ccggaatatt aatagatcat ggagataatt
     1981 aaaatgataa ccatctcgca aataaataag tattttactg ttttcgtaac agttttgtaa
     2041 taaaaaaacc tataaatatt ccggattatt cataccgtcc caccatcggg cgcggatccc
     2101 gatgagcggc ctgaacgata tttttgaagc gcagaaaatt gaatggcatg aaggcagcgc
     2161 tggaggttca ggtgggactc aagagcagga tattgaagcc gccagttcgc agcggttaag
     2221 agtgctctat agaattgaaa agatgtttct ccagctttta gagattgagg agggctggaa
     2281 ataccgccct cccccaccgt gttttagcga acaacagtcc aaccaagtcg aaaaactttt
     2341 ccaaacccta aaaacacagg agcagaataa cctggaggaa gcggctgacg gtttcttaca
     2401 ggttctaagt gtcaggaaag gtaaggcgct cgtagcgcgg ctattgccgt ttcttccaca
     2461 agaccaagca gtcacaatac tcttagctat cactcaccac ctcccactat tggtgcggag
     2521 ggacgttgca gaccaagctc tgcaaatgtt gtttaagccc cttggtaaat gcatttccca
     2581 cctcacgctc catgagcttc tgcaaggact ccaaggattg acacttttgc cccctggatc
     2641 atctgaacga cccgtgacag tcgttttgca gaatcagttt ggaattagtc tactgtacgc
     2701 acttttatca catggggagc aattggtgag cttgcactct tcactggaag agcccaacag
     2761 cgaccacacg gcatggaccg atatggtagt actgatcgct tgggagatag cgcaaatgcc
     2821 tacagcttcg cttgctgagc ctctggcatt tcctagtaac ctgttgcccc tattctgtca
     2881 ccatgtcgat aaacaattag tccagcagct agaggcacga atggagtttg cttggatcta
     2941 tggcggaagt ggacatcacc accatcatca ctgatgaagc ttgtcgagaa gtactagagg
     3001 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3061 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3121 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3181 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3241 tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat
     3301 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     3361 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     3421 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc
     3481 aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt
     3541 ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag
     3601 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3661 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3721 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3781 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3841 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3901 cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3961 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4021 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4081 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4141 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4201 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4261 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4321 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4381 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4441 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4501 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4561 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4621 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4681 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4741 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4801 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4861 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4921 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4981 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5041 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5101 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5161 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5221 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5281 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5341 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5401 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5461 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5521 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5581 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5641 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5701 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5761 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5821 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5881 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5941 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6001 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     6061 accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat gccctgcgta
     6121 agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa actatgacaa
     6181 taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta tgctgtgaaa
     6241 aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa gtgcaaattg
     6301 cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt cgcggcgttg
     6361 tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta
     6421 ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt
     6481 atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc
     6541 aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg
     6601 gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct gctcaaactt
     6661 gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc
     6721 gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga
     6781 gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt
     6841 gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt
     6901 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctcca
//
