LOCUS       Exported                4757 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses thermostable variant of human carbonic anhydrase II with 
            TAG stop codon at site 186 and C-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-CA TAG186
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4757)
  TITLE     Bacterial expression plasmids for genetic code expansion laboratory 
            course
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4757)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4757
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        13..297
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     220..239
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1111..1128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1170..1187)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1340..1426
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1518..1545
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1564..1655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1656..2516
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1874..1893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2687..3275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3176..3195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3429..3446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3461..3601
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3527..3546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3687..3709
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3865..4743)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
       61 tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
      121 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
      181 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
      241 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
      301 cgttttttgg gctaacagga ggaattaacc atggcgcatc attggggtta cggtaaacac
      361 aacggtccgg agcattggca caaagatttt ccaattgcga agggcgaacg tcaaagcccg
      421 gttgacattg atacgcacac ggcaaagtac gacccgagcc tgaaaccgct gagcgtttcc
      481 tatgaccagg ctacgagcct gcgtatcctg aacaatggcc acaccttcaa cgtggagttt
      541 gatgattccc aagataaggc ggttctgaaa ggtggtccgt tggatggcac ctaccgcctg
      601 atccaatttc actttcactg gggtagccac gacggtcagg gcagcgagca taccgtggac
      661 aaaaagaagt atgcagccga actgcacctg gtgcattgga acacgaagta cggcgacttc
      721 ggtaaagcgg tccagcaacc ggacggtctg gctgttctgg gtattttcct gaaggtcggc
      781 agcgcgaacc cgggtctgca gaaagtggtt gacgtgttgg actctatcaa gaccaaaggc
      841 aagagcgcgg acttcaccaa tttcgatccg cgtggtctgc tgccgtagag cctggattac
      901 tggacttatc cgggcagcct gaccaccccg ccattgctgg agtgcgtgac ctggatcgtc
      961 ttgaaagaac cgatcagcgt tagctctgaa caggtcagca agttccgcaa gctgaatttc
     1021 aatggtgagg gcgagccgga agaaccgatg gtcgataatt ggcgtcctac ccaaccgctg
     1081 aaaaaccgcc agattaaagc atcctttaag catcaccacc accatcacta aaagcttggc
     1141 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     1201 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1261 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     1321 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1381 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     1441 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     1501 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     1561 tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1621 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1681 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1741 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1801 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1861 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     1921 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1981 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2041 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2101 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2161 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2221 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2281 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2341 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2401 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2461 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2521 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2581 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2641 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2761 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2821 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2881 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2941 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3001 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3061 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3121 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3181 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3241 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3301 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3361 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3421 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3481 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     3541 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     3601 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3661 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3721 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata
     3781 atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc
     3841 gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc
     3901 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
     3961 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
     4021 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
     4081 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
     4141 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
     4201 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
     4261 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
     4321 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
     4381 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
     4441 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
     4501 gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg
     4561 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
     4621 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
     4681 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
     4741 catacttttc atactcc
//
