LOCUS       Exported                9364 bp ds-DNA     circular SYN 30-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9364)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 21, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9364
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    442..1124
                     /label=COL E1 ORIGIN OF REPLICATION)
     misc_feature    1260..1277
                     /label=L4440
     misc_feature    1374..1394
                     /label=LAC O3
     misc_feature    1406..1486
                     /label=LAC PROMOTER
     misc_feature    1469..1485
                     /label=LAC OPERATOR (Lutz & Bujard paper)
                     
     misc_feature    1492..1512
                     /label=M13-rev
     misc_feature    1530..1549
                     /label=T3
     misc_feature    1803..1983
                     /label=5' LTR (truncated) SnapGene
     misc_feature    2027..2152
                     /label=HIV-1 PSI PACKAGING SIGNAL (SnapGene)
     misc_feature    2645..2878
                     /label=REV RESPONSE ELEMENT RRE (SnapGene)
     misc_feature    3374..3491
                     /label=cPPT HIV TRACT (SnapGene)
     primer_bind     complement(3483..3512)
                     /label=Rev_BB_DeleteNeo
     primer_bind     3483..3512
                     /label=Fwd_DeleteNeo
     enhancer        3513..3816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3817..4020
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    4043..4123
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    4125..4152
                     /label=KOZAK SEQUENCE
     primer_bind     complement(4152..4185)
                     /label=Rev_BB_51
     misc_feature    4153..4176
                     /label=Flag Tag
     primer_bind     4167..4198
                     /label=144
     CDS             4177..4884
                     /codon_start=1
                     /label=uTEV3 protease
                     /label=TEV protease
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.6"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGTFWKHWIQTKDGQCGNPLVSTRDGFI
                     VGIHSASNFTNTNNYFASVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     primer_bind     complement(4863..4893)
                     /label=Rev_BB_53
     primer_bind     4873..4906
                     /label=Fwd_53
     misc_feature    4885..5358
                     /label=ecDHFR_C12 (no start)
                     /label=ecDHFR (no start)
     primer_bind     4902..4946
                     /label=Fwd_H12RT18NV18A
     primer_bind     complement(4902..4935)
                     /label=Rev_H12RT18NV18A
     misc_feature    4915..4917
                     /label=H12R
     misc_feature    4933..4935
                     /label=T18N
     misc_feature    4936..4938
                     /label=V19A
     primer_bind     5070..5092
                     /label=Fwd_S67G
     primer_bind     complement(5070..5092)
                     /label=Rev_S67G
     misc_feature    5080..5082
                     /label=S67G
     primer_bind     5088..5115
                     /label=Fwd_W74R
     primer_bind     complement(5089..5119)
                     /label=Rev_W74R
     misc_feature    5101..5103
                     /label=W74R
     primer_bind     5203..5249
                     /label=T113S_E120D_Fwd
     primer_bind     complement(5203..5231)
                     /label=T113S_E120D_Rev
     misc_feature    5218..5220
                     /label=E113S
     misc_feature    5239..5241
                     /label=E120D
     primer_bind     5301..5328
                     /label=Q146L_Fwd
     primer_bind     complement(5306..5336)
                     /label=Q146L_Rev
     misc_feature    5317..5319
                     /label=Q146L
     primer_bind     5341..5363
                     /label=FWD_BB_53
     primer_bind     complement(5341..5363)
                     /label=Rev_53
     misc_feature    5362..5948
                     /label=IRES2 (from SnapGene pIRES2EGFPconstruct)
     misc_feature    5949..6659
                     /locus_tag="mCherry"
                     /label=mCherry
     primer_bind     complement(6672..6700)
                     /label=Rev_mcherry_48
     primer_bind     6686..6716
                     /label=Fwd_48_BB
     misc_feature    6739..7327
                     /label=WPRE (SnapGene)
     misc_feature    7460..7693
                     /label=3' LTR (SnapGene)
     misc_feature    7765..7886
                     /label=SV40 POLY(A)
     misc_feature    7765..7886
                     /label=SV40 POLY(A) (SnapGene)
     misc_feature    7905..8040
                     /label=SV40 ORI (SnapGene)
     misc_feature    8072..8091
                     /label=T7 REV. SEQ. PRIMER)
     misc_feature    8073..8091
                     /label=T7 PROMOTER
     misc_feature    8101..8118
                     /label=M13-fwd
     misc_feature    8189..8257
                     /label=LAC Z ALPHA FRAGMENT
     misc_feature    8261..8536
                     /label=F1 INTERGENIC REGION
     misc_feature    8746..8850
                     /label=AmpR Promoter
     misc_feature    8781..8809
                     /label=PROMOTER BETA LACTAMASE
     primer_bind     complement(9215..9245)
                     /label=REV_Split_pJY0
     primer_bind     9227..9252
                     /label=FWD_Split_pJY0
ORIGIN
        1 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
       61 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
      121 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      181 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      241 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
      301 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
      361 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
      421 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
      481 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      541 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      601 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      661 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      721 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      781 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      841 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      901 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      961 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1021 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1081 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1141 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1201 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1261 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     1321 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     1381 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     1441 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     1501 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     1561 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     1621 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     1681 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     1741 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacaataa
     1801 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     1861 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     1921 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1981 gcagtggcgc ccgaacaggg acctgaaagc gaaagggaaa ccagagctct ctcgacgcag
     2041 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     2101 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     2161 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     2221 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     2281 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     2341 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     2401 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     2461 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     2521 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     2581 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     2641 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcct
     2701 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     2761 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     2821 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     2881 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     2941 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     3001 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     3061 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     3121 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     3181 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     3241 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     3301 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     3361 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     3421 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     3481 ttcaaaattt tatcgatcac gagactagcc tccgttacat aacttacggt aaatggcccg
     3541 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     3601 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     3661 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     3721 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     3781 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     3841 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     3901 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3961 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     4021 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
     4081 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcgagctc ggatccagta
     4141 cccttcacca tggactacaa ggacgacgac gacaagggag aaagcttgtt taaggggccg
     4201 cgtgattaca acccgatatc gagcaccatt tgtcatttga cgaatgaatc tgatgggcac
     4261 acaacatcgt tgtatggtat tggatttggt cccttcatca ttacaaacaa gcacttgttt
     4321 agaagaaata atggaacact gttggtccaa tcactacatg gtgtattcaa ggtcaagaac
     4381 accacgactt tgcaacaaca cctcattgat gggagggaca tgataattat tcgcatgcct
     4441 aaggatttcc caccatttcc tcaaaagctg aaatttagag agccacaaag ggaagagcgc
     4501 atatgtcttg tgacaaccaa cttccaaact aagagcatgt ctagcatggt gtcagacact
     4561 agttgcacat tcccttcatc tgatggcaca ttctggaagc attggattca aaccaaggat
     4621 gggcagtgtg gcaatccatt agtatcaact agagatgggt tcattgttgg tatacactca
     4681 gcatcgaatt tcaccaacac aaacaattat ttcgcaagcg tgccgaaaaa cttcatggaa
     4741 ttgttgacaa atcaggaggc gcagcagtgg gttagtggtt ggcgattaaa tgctgactca
     4801 gtattgtggg ggggccataa agttttcatg gtgaaacctg aagagccttt tcagccagtt
     4861 aaggaagcga ctcaactcat gaatatcagt ctgattgcgg cgttagcggt agatcgcgtt
     4921 atcggcatgg aaaacgcgat gccgtggaac ctgcctgccg atctcgcctg gtttaaacgc
     4981 aacaccttaa ataaacccgt gattatgggc cgccatacct gggaatcaat cggtcgtccg
     5041 ttgccaggac gcaaaaatat tatcctcagc agtcaaccgg gcacggacga tcgcgtaacg
     5101 agagtgaagt cggtggatga agccatcgcg gcgtgtggtg acgtaccaga aatcatggtt
     5161 attggcggcg gtcgcgttta tgaacagttc ttgccaaaag cgcaaaaact gtatctgagc
     5221 catatcgacg cagaagtgga tggcgacacc catttcccgg attacgagcc ggatgactgg
     5281 gaatcggtat tcagcgaatt ccacgatgct gatgcgctta actctcacag ctattgcttt
     5341 gagattctgg agcggcgatg acccctctcc ctcccccccc cctaacgtta ctggccgaag
     5401 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     5461 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     5521 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     5581 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     5641 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     5701 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     5761 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     5821 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     5881 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     5941 ccacaaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     6001 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     6061 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     6121 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     6181 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     6241 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     6301 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     6361 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     6421 aggacggcgc cctgaagggc gagatcaggc agaggctgaa gctgaaggac ggcggccact
     6481 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     6541 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     6601 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaga
     6661 gcggccgcac tcgagatatc tagacccagc tttcttgtac aaagtggttg atatccagca
     6721 cagtggcggc cgctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6781 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6841 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6901 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6961 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     7021 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     7081 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     7141 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     7201 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     7261 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     7321 tccccgcctg gaattctgca gatatccgga ctagtgatct aattctaccg gggatccgtc
     7381 gagggcccgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     7441 ttaaaagaaa aggggggact ggaagggcta gctcactccc aacgaagaca agatctgctt
     7501 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctgc
     7561 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7621 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7681 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7741 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     7801 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7861 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     7921 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7981 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     8041 taggcttttg cgtcgagacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     8101 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     8161 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     8221 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc gacgcgccct gtagcggcgc
     8281 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     8341 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     8401 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     8461 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     8521 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     8581 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     8641 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     8701 attaacgttt acaatttccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     8761 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     8821 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     8881 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     8941 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     9001 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     9061 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     9121 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     9181 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     9241 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     9301 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     9361 atac
//
