LOCUS       Exported                5179 bp ds-DNA     circular SYN 01-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 1, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1..5179
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          4669..5148
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label= [Split]
                     /note=" [Split]"
     misc_feature    34..87
                     /label=Tet promoter
                     /note="""/ApEinfo_fwdcolor=#00ff00"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=green"
     CDS             complement(100..113)
                     /codon_start=1
                     /label=oTN307 - reverse sequencing primer
                     /note="oTN307 - reverse sequencing primer"
                     /translation="EFGQ"
     misc_feature    117..128
                     /label=RBS
                     /note="""/ApEinfo_fwdcolor=#ff00ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=green"
     CDS             149..943
                     /codon_start=1
                     /label=E. coli R166Q TYMS
                     /note="E. coli R166Q TYMS"
                     /note="/ApEinfo_fwdcolor=#ff0000"
                     /note="/ApEinfo_revcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /translation="MKQYLELMQKVLDEGTQKNDRTGTGTLSIFGHQMRFNLQDGFPLV
                     TTKRCHLRSIIHELLWFLQGDTNIAYLHENNVTIWDEWADENGDLGPVYGKQWRAWPTP
                     DGRHIDQITTVLNQLKNDPDSRRIIVSAWNVGELDKMALAPCHAFFQFYVADGKLSCQL
                     YQQSCDVFLGLPFNIASYALLVHMMAQQCDLEVGDFVWTGGDTHLYSNHMDQTHLQLSR
                     EPRPLPKLIIKRKPESIFDYRFEDFEIEGYDPHPGIKAPVAI"
     protein_bind    956..1000
                     /label=S-Tag
                     /note="S-Tag"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     primer_bind     complement(1038..1056)
                     /label=T7 Terminator Primer
                     /note="T7 Terminator Primer"
                     /note="""/label=T7 Terminator Primer"
                     /note="/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     terminator      1052..1099
                     /label=T7 Terminator
                     /note="T7 Terminator"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     CDS             complement(1319..1978)
                     /codon_start=1
                     /label=CAT
                     /note="CAT"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      2340..3252
                     /label=P15A ori
                     /note="P15A ori"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     CDS             complement(3376..4458)
                     /codon_start=1
                     /label=lacI gene
                     /note="lacI gene"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4539..4558
                     /label=pACYCDuetUP1 Primer
                     /note="pACYCDuetUP1 Primer"
                     /note="""/label=pACYCDuetUP1 Primer"
                     /note="/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     promoter        4582..4598
                     /label=T7 Promoter
                     /note="T7 Promoter"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     protein_bind    4603..4623
                     /label=lacO;
                     /note="lacO;"
                     /note="""/ApEinfo_fwdcolor=pink"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=pink"
     misc_feature    4656..4661
                     /label=RBS-1
                     /note="""/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 6 offset 0"""
                     /note="/ApEinfo_revcolor=green"
     CDS             4669..5148
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=M42F/G121V DHFR E. coli
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIFGRH
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEVDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     misc_feature    5149..5153
                     /label=BAR DHFR
                     /note="BAR DHFR"
                     /note="""/ApEinfo_fwdcolor=#ffb900"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=green"
     misc_feature    5174..5179
                     /label=BAR ThyS
                     /note="BAR ThyS"
                     /note="""/ApEinfo_fwdcolor=#ffb900"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"""
                     /note="/ApEinfo_revcolor=green"
ORIGIN
        1 cttaagtcga acagaaagta atcgtattgt acatccctat cagtgataga gattgacatc
       61 cctatcagtg atagagatac tgagcacatc agcaggacgc actgaccgaa ttcattaaag
      121 aggagaaagg taccatgctg cagatctcat gaaacagtac ctggagctga tgcaaaaagt
      181 tctggatgag ggtacccaga aaaacgaccg cacgggtacc ggaacgctga gcatttttgg
      241 ccatcagatg cgttttaacc tgcaggacgg attcccgctg gttacgacca aacgctgcca
      301 cctgcgtagc attattcatg agctgctgtg gtttctgcaa ggtgacacta acatcgcgta
      361 tctgcacgaa aacaatgtga cgatctggga tgaatgggcc gatgaaaacg gcgatctggg
      421 cccagtgtat ggtaaacagt ggcgcgcctg gccaacgcca gatggccgcc acatcgatca
      481 gatcacgacc gtgctgaatc aactgaaaaa cgacccggac agccgccgca ttattgtttc
      541 cgcgtggaat gtgggtgaac tggataaaat ggcgctggcg ccgtgccatg cattcttcca
      601 gttttacgtt gcggacggta agctgagctg tcaactttat cagcagagct gcgatgtttt
      661 cctcggcctg ccgttcaata tcgccagcta cgcgttactg gtgcatatga tggcgcagca
      721 gtgcgacctg gaggtgggtg attttgtctg gaccggcggt gatacccacc tgtacagcaa
      781 ccatatggac caaacgcatc tgcaattgag ccgtgagccg cgtccgctgc cgaaactgat
      841 tattaagcgc aaacccgaga gcatctttga ctatcgtttc gaagacttcg agattgaagg
      901 ctacgatccg cacccgggta tcaaagcgcc ggtggctatc taactcgagt ctggtaaaga
      961 aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt
     1021 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1081 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct
     1141 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     1201 ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta
     1261 tcacttattc aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaatt
     1321 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     1381 ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     1441 cttgcgtata atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc atattggcca
     1501 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     1561 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     1621 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     1681 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     1741 cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa
     1801 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     1861 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     1921 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     1981 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     2041 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     2101 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     2161 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac
     2221 tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt
     2281 ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag
     2341 cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc
     2401 ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat
     2461 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga
     2521 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga
     2581 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac
     2641 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg
     2701 tttcccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     2761 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     2821 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     2881 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     2941 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     3001 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     3061 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     3121 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     3181 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     3241 atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg
     3301 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     3361 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3421 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     3481 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     3541 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     3601 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc
     3661 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     3721 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     3781 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     3841 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     3901 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     3961 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     4021 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     4081 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     4141 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     4201 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     4261 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     4321 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     4381 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     4441 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     4501 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     4561 gcgactcctg cattaggaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     4621 attcccctgt agaaataatt ttgtttaact ttaataagga gatataccat gatcagtctg
     4681 attgcggcgt tagcggtaga tcgcgttatc ggcatggaaa acgccatgcc gtggaacctg
     4741 cctgccgatc tcgcctggtt taaacgcaac accttaaata aacccgtgat tttcggccgc
     4801 catacctggg aatcaatcgg tcgtccgttg ccaggacgca aaaatattat cctcagcagt
     4861 caaccgggta cggacgatcg cgtaacgtgg gtgaagtcgg tggatgaagc catcgcggcg
     4921 tgtggtgacg taccagaaat catggtgatt ggcggcggtc gcgtttatga acagttcttg
     4981 ccaaaagcgc aaaaactgta tctgacgcat atcgacgcag aagtggaagt cgacacccat
     5041 ttcccggatt acgagccgga tgactgggaa tcggtattca gcgaattcca cgatgctgat
     5101 gcgcagaact ctcacagcta ttgctttgag attctggagc ggcggtaaca ggcgtcgaca
     5161 agcttgcggc cgcataatg
//
