LOCUS       Exported                3923 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pOPINCDE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3923)
  TITLE     OPPF-UK pOPIN vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3923
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    168..189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        204..234
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    242..258
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     247..269
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     266..282
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     266..282
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             278..451
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(298..315)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(298..314)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(307..329)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             471..488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             508..552
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(590..608)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      604..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(822..1613)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1614..1705)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1753..2491)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2701..3783)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3745..3764
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3784..3861)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3907..3
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3907..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcacacc atcaccacca tcacagcagc ggtctggaag ttctgtttca
      121 gggtaccgtc gacccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      181 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      241 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
      301 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      361 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      421 ttcccaacag ttgcgcagcc tgaatggcga atggcgccaa gctgtttaaa catcaccatc
      481 accatcacta agcttctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag
      541 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
      601 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct
      661 caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag
      721 caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg
      781 ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg
      841 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
      901 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
      961 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     1021 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     1081 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     1141 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     1201 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     1261 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     1321 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     1381 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     1441 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     1501 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     1561 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc
     1621 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1681 gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg
     1741 agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag
     1801 gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc
     1861 gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga
     1921 ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg
     1981 tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac
     2041 ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta
     2101 tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca
     2161 gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg
     2221 taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta
     2281 aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg
     2341 gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc
     2401 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag
     2461 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt
     2521 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     2581 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     2641 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     2701 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2761 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     2821 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     2881 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     2941 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     3001 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     3061 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     3121 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     3181 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     3241 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     3301 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     3361 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     3421 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     3481 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     3541 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     3601 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     3661 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     3721 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     3781 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     3841 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     3901 ggaaattaat acgactcact ata
//
