LOCUS       pVY-17:_TC-TEM1-tag3    3994 bp ds-DNA     circular     05-OCT-2022
DEFINITION  .
FEATURES             Location/Qualifiers
     rep_origin      complement(39..777)
                     /label="cloDF13"
     primer          413..432
                     /label="VY90"
                     /note="sequence: ccaagctgggctgtaagcaa"
     modified_base   852..866
                     /label="USER junction"
     terminator      867..911
                     /label="rrnB1 transcriptional terminator"
     modified_base   912..926
                     /label="USER junction"
     misc_feature    927..967
     CDS             complement(927..1805)
                     /label="araC"
     misc_feature    978..978
                     /label="originally an 'a' in annotation"
     misc_feature    1017..1017
                     /label="originally a t in annotation"
     misc_feature    1083..1083
                     /label="originally an a in annotation"
     misc_feature    1287..1287
                     /label="originally a c in annotation"
     misc_feature    1452..1452
                     /label="originally an a in annotation"
     misc_feature    1611..1611
                     /label="originally a g in annotation"
     primer          1673..1692
                     /label="VY93"
                     /note="sequence: aacctttcattcccagcggt"
     misc_feature    1758..1758
                     /label="originally a g in annotation"
     primer          complement(1787..1815)
                     /label="VY97"
                     /note="sequence: tatgaaaagtatggctgaagcgcaaaatg"
     source          1806..2171
                     /label="source - from pBAD mOrange"
     promoter        1832..2116
                     /label="araBAD promoter"
     primer_bind     2039..2058
                     /label="pBAD Forward"
     primer          complement(2135..2172)
                     /label="VY145"
                     /note="sequence: ATGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTC"
     RBS             2143..2165
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
     primer          2153..2224
                     /label="VY148"
                     /note="sequence: tttaagaaggagatatacatATGGGTTGTTGCCCTGGGTGTTGTGGTGGCGCTAGTATTCAACATTTCCGTG"
     gene            2173..2205
                     /label="TC-FlAsH Tag"
     CDS             2173..2205
                     /label="Translation 2173-2205"
                     /translation="MGCCPGCCGGA"
     misc_feature    2179..2196
                     /label="tetracysteine tag"
     gene            2206..3144
                     /label="TEM1-tag3"
     CDS             2206..3144
                     /label="Translation 2206-3144"
                     /translation="SIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHWGGSKKRKKRGGSKKRKKRGGSKKRKKR*"
     primer          complement(3042..3080)
                     /label="VY147"
                     /note="sequence: TTGCGTTTCTTGCTTCCTCCCCAATGCTTAATCAGTGAG"
     primer          3061..3078
                     /label="VY144"
                     /note="sequence: GGAGGAAGCAAGAAACGC"
     misc_feature    3061..3141
                     /label="tag3"
     modified_base   3148..3154
                     /label="USER junction"
     primer          3148..3172
                     /label="VY75"
                     /note="sequence: acttaattaacggcactcctcagcc"
     modified_base   3155..3164
                     /label="USER junction"
     terminator      complement(3172..3207)
                     /label="termination of cat transcript is slightly weakerthan in opposite direction"
     modified_base   3208..3220
                     /label="USER junction"
     CDS             complement(3221..3880)
                     /label="CAT/CamR variant from pACYCDuet-1"
     modified_base   3296..3296
                     /label="annotated as a G in pACYCDuet cat markerannotation, here it is an A, but this mutation is silentfrom codon GTC (Val, 25% codon usage) to GTT (Val, 21%usage), so it should not be of functional relevance"
     primer          3793..3814
                     /label="VY89"
                     /note="sequence: gagTaactgactgaaatgcctc"
     promoter        3881..3978
                     /label="from pACYCDuet-1"
     modified_base   3979..3994
                     /label="USER junction"
ORIGIN
        1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
       61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
      121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
      181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
      241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
      301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
      361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
      421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
      481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
      541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
      601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
      661 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
      721 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
      841 agcccaggag gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tagacttagg gaccctttat gacaacttga cggctacatc attcactttt
      961 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     1021 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     1081 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     1141 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     1201 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     1261 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     1321 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     1381 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     1441 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     1501 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     1561 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     1621 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     1681 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     1741 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     1801 gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga
     1861 cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt
     1921 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     1981 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
     2041 gccatagcat ttttatccat aagattagcg gatactacct gacgcttttt atcgcaactc
     2101 tctactgttt ctccataccc gtttttttgg gctagaaata attttgttta actttaagaa
     2161 ggagatatac atATGGGTTG TTGCCCTGGG TGTTGTGGTG GCGCTAGTAT TCAACATTTC
     2221 CGTGTCGCCC TTATTCCCTT TTTTGCGGCA TTTTGCCTTC CTGTTTTTGC TCACCCAGAA
     2281 ACGCTGGTGA AAGTAAAAGA TGCTGAAGAT CAGTTGGGTG CACGAGTGGG TTACATCGAA
     2341 CTGGATCTCA ACAGCGGTAA GATCCTTGAG AGTTTTCGCC CCGAAGAACG TTTTCCAATG
     2401 ATGAGCACTT TTAAAGTTCT GCTATGTGGC GCGGTATTAT CCCGTATTGA CGCCGGGCAA
     2461 GAGCAACTCG GTCGCCGCAT ACACTATTCT CAGAATGACT TGGTTGAGTA CTCACCAGTC
     2521 ACAGAAAAGC ATCTTACGGA TGGCATGACA GTAAGAGAAT TATGCAGTGC TGCCATAACC
     2581 ATGAGTGATA ACACTGCGGC CAACTTACTT CTGACAACGA TCGGAGGACC GAAGGAGCTA
     2641 ACCGCTTTTT TGCACAACAT GGGGGATCAT GTAACTCGCC TTGATCGTTG GGAACCGGAG
     2701 CTGAATGAAG CCATACCAAA CGACGAGCGT GACACCACGA TGCCTGTAGC AATGGCAACA
     2761 ACGTTGCGCA AACTATTAAC TGGCGAACTA CTTACTCTAG CTTCCCGGCA ACAATTAATA
     2821 GACTGGATGG AGGCGGATAA AGTTGCAGGA CCACTTCTGC GCTCGGCCCT TCCGGCTGGC
     2881 TGGTTTATTG CTGATAAATC TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT CATTGCAGCA
     2941 CTGGGGCCAG ATGGTAAGCC CTCCCGTATC GTAGTTATCT ACACGACGGG GAGTCAGGCA
     3001 ACTATGGATG AACGAAATAG ACAGATCGCT GAGATAGGTG CCTCACTGAT TAAGCATTGG
     3061 GGAGGAAGCA AGAAACGCAA GAAGCGTGGC GGTAGCAAAA AGCGTAAGAA GCGTGGAGGT
     3121 TCAAAGAAAC GTAAAAAGCG ATAAtaaact taattaacgg cactcctcag ccaagtcaaa
     3181 agcctccgac cggaggcttt tgactacatg cccatggcgt ttacgccccg ccctgccact
     3241 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     3301 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     3361 ccatagtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     3421 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     3481 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     3541 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     3601 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     3661 ggaactccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     3721 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     3781 tataggtaca ttgagTaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     3841 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     3901 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     3961 tggaacctct tacgtgccaA gccaaatagg ccgt
//