LOCUS       Exported                4757 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses thermostable variant of human carbonic anhydrase II with 
            TAG stop codon at site 20 and C-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-CA TAG20
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  JOURNAL   Unpublished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 12, 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        23..307
                     /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     230..249
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1121..1138
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1180..1197)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1350..1436
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1528..1555
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1574..1665
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1666..2526
                     /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(1884..1903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2697..3285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3186..3205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3439..3456
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3471..3611
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3537..3556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3697..3719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3875..4753)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
       61 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
      121 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat
      181 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca
      241 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt
      301 tctccatacc cgttttttgg gctaacagga ggaattaacc atggcgcatc attggggtta
      361 cggtaaacac aacggtccgg agcattggca caaagattag ccaattgcga agggcgaacg
      421 tcaaagcccg gttgacattg atacgcacac ggcaaagtac gacccgagcc tgaaaccgct
      481 gagcgtttcc tatgaccagg ctacgagcct gcgtatcctg aacaatggcc acaccttcaa
      541 cgtggagttt gatgattccc aagataaggc ggttctgaaa ggtggtccgt tggatggcac
      601 ctaccgcctg atccaatttc actttcactg gggtagccac gacggtcagg gcagcgagca
      661 taccgtggac aaaaagaagt atgcagccga actgcacctg gtgcattgga acacgaagta
      721 cggcgacttc ggtaaagcgg tccagcaacc ggacggtctg gctgttctgg gtattttcct
      781 gaaggtcggc agcgcgaacc cgggtctgca gaaagtggtt gacgtgttgg actctatcaa
      841 gaccaaaggc aagagcgcgg acttcaccaa tttcgatccg cgtggtctgc tgccggagag
      901 cctggattac tggacttatc cgggcagcct gaccaccccg ccattgctgg agtgcgtgac
      961 ctggatcgtc ttgaaagaac cgatcagcgt tagctctgaa caggtcagca agttccgcaa
     1021 gctgaatttc aatggtgagg gcgagccgga agaaccgatg gtcgataatt ggcgtcctac
     1081 ccaaccgctg aaaaaccgcc agattaaagc atcctttaag catcaccacc accatcacta
     1141 aaagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     1201 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     1261 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     1321 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1381 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1441 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1501 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     1561 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1621 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1681 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1741 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1801 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1861 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1921 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1981 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2041 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2101 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2161 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2221 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2281 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2341 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2401 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2461 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2521 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2581 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2641 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2701 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2761 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2821 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2881 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2941 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3001 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3061 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3121 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3181 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3241 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3301 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3361 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3421 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3481 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3541 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3601 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3721 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc
     3781 ggcatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
     3841 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
     3901 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
     3961 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
     4021 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
     4081 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
     4141 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
     4201 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
     4261 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
     4321 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
     4381 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
     4441 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
     4501 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
     4561 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
     4621 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     4681 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     4741 gcgcttcagc catactt
//
