LOCUS       pACYCDuet-1_ChrA        4181 bp DNA     circular SYN 27-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4181)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jul 27, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             71..316
                     /codon_start=1
                     /label=ChrA-C63S
                     /translation="MGSSHHHHHHSQMKIPALVMASLLAVSVSGQTTKPVKKGTKSVKK
                     VKKMDSPKTVKAEPTKVVKRDTILKHGGGSPACGMG"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    293..295
                     /locus_tag="C63"
                     /label=C63S
                     /label=C63
     promoter        387..405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    406..430
                     /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             539..583
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      635..682
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(902..1561)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1562..1664)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(2190..2734)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     CDS             complement(2959..4041)
                     /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"
     promoter        complement(4042..4119)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4165..4181,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccagatga agattccggc
      121 tctggtgatg gccagcctgc tggcggtatc agtgagcggg caaaccacga aaccggtcaa
      181 aaaaggcacg aaaagtgtta aaaaagtgaa aaaaatggat agcccaaaaa ccgttaaagc
      241 agaacccacc aaagtcgtta aacgtgacac gatcctgaaa catggcggcg gttctccggc
      301 gtgcggtatg gggtgaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
      361 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
      421 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatggcaga
      481 tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa
      541 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
      601 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      661 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
      721 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
      781 ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta
      841 ttatcactta ttcaggcgta gcaccaggcg tttaagggca ccaataactg ccttaaaaaa
      901 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
      961 catggaagcc atcacagacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     1021 cgccttgcgt ataatatttg cccatagtga aaacgggggc gaagaagttg tccatattgg
     1081 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     1141 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     1201 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     1261 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     1321 caccgtcttt cattgccata cggaactccg gatgagcatt catcaggcgg gcaagaatgt
     1381 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     1441 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     1501 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     1561 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     1621 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
     1681 aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga
     1741 agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact
     1801 tactgattta gtgtatgatg gtgtttttga ggtgctccag tggcttctgt ttctatcagc
     1861 tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat
     1921 cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     1981 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     2041 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2101 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     2161 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     2221 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     2281 gcgtttcccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     2341 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     2401 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     2461 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     2521 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     2581 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     2641 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     2701 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     2761 atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     2821 acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact
     2881 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     2941 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3001 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     3061 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     3121 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     3181 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     3241 gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     3301 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     3361 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     3421 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     3481 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     3541 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     3601 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     3661 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     3721 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     3781 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     3841 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     3901 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     3961 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     4021 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     4081 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     4141 tatgcgactc ctgcattagg aaattaatac gactcactat a
//
