LOCUS       Exported                8386 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAM3110
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8386)
  AUTHORS   Ditty JL, Canales SR, Anderson BE, Williams SB, Golden SS
  TITLE     Stability of the Synechococcus elongatus PCC 7942 circadian clock 
            under directed anti-phase expression of the kai genes.
  JOURNAL   Microbiology. 2005 Aug;151(Pt 8):2605-13.
  PUBMED    16079339
REFERENCE   2  (bases 1 to 8386)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8386
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      725..1313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1214..1233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1467..1484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1499..1639
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1565..1584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1725..1747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             3558..4349
                     /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"
     primer_bind     4782..4804
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4796..4813
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4797..4813
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4830..5061
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     CDS             complement(5457..5762)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(5559..5578)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6107..6766)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6681..6700
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6767..6869)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(7014..7245)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     primer_bind     complement(7263..7282)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7264..7282)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
       61 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
      241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      421 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      841 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
      901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1441 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1501 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     1561 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     1621 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1741 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag gatcgatccg gcagccggcg
     1801 gagcgctgct ttcttggcaa gcggtcgcca gccccaacgc cagggctgcc agcccgaaac
     1861 agcggggcaa ggcagcttgg aagggcgatc gcagcacggg catggcaatg tctctctgaa
     1921 ggaatgcaga ccttattcgt acagccaggg ttgaatcgtg ggggtccaat cacttagctc
     1981 tgctgggcta aaccagagag caatttcctg ttgtgctgtt tcgattgcat ccgagccatg
     2041 gatgatgttg cggccaatat tgacaccaaa atcaccacgg atggtgcccg gttctgccgt
     2101 cagcggattg gtagcgccga tcaacttgcg agcagccgcc acaacgcctt cgccttccaa
     2161 gacgatcgcc acgatcggcc cagaggtgat gaactcgacg aggccattga agaaggggcg
     2221 ctcgcggtgg acagcatagt gctgttcggc cagctcgcga ctgggcttca gctgctttag
     2281 gcccaccagt ttgaagcctt tttgctcaaa gcggccgatg atcgtaccga ccaaaccccg
     2341 ctgaacgcca tcgggcttga tggcaataaa tgtgcgttcc acagacatct agatagtcct
     2401 caagacgagg caagcattga gcttgccttc ctatggttcg ggatcactgg gattcttgac
     2461 aagcgatcgc ggtcacatcg ctatctctta ggacttcgca gcgggcgagt cggattgacc
     2521 cggtagggat ttcgccagat caatgcccgt ggtttgtttc agcttctcca gcaagctagc
     2581 gatttgggta gcgctgcctt ccccttcgcc aatcacagtg atcgactcca cgtcgatatc
     2641 tggcacggtg cctgaaagcg tgacgagcag ggactcgaag cttgcatgcc tgcaggtcga
     2701 ctctagagct ttatgcttgt aaaccgtttt gtgaaaaaat ttttaaaata aaaaagggga
     2761 cctctagggt ccccaattaa ttagtaatat aatctattaa aggtcattca aaaggtcatc
     2821 caccggatca attcccctgc tcgcgcaggc tgggtgccaa gctctcgggt aacatcaagg
     2881 cccgatcctt ggagcccttg ccctcccgca cgatgatcgt gccgtgatcg aaatccagat
     2941 ccttgacccg cagttgcaaa ccctcactga tccgcatgcc cgttccatac agaagctggg
     3001 cgaacaaacg atgctcgcct tccagaaaac cgaggatgcg aaccacttca tccggggtca
     3061 gcaccaccgg caagcgccgc gacggccgag gtcttccgat ctcctgaagc cagggcagat
     3121 ccgtgcacag caccttgccg tagaagaaca gcaaggccgc caatgcctga cgatgcgtgg
     3181 agaccgaaac cttgcgctcg ttcgccagcc aggacagaaa tgcctcgact tcgctgctgc
     3241 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     3301 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     3361 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     3421 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     3481 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     3541 aaacaaagtt aaacatcatg agggaagcgg tgatcgccga agtatcgact caactatcag
     3601 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     3661 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     3721 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     3781 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     3841 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     3901 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     3961 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     4021 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     4081 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     4141 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     4201 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     4261 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     4321 gcgagatcac caaggtagtc ggcaaataat gtctaacaat tcgttcaagc cgacgccgct
     4381 tcgcggcgcg gcttaactca agcgttagat gcactaagca cataattgct cacagccaaa
     4441 ctatcaggtc aagtctgctt ttattatttt taagcgtgca taataagccc tacacaaatt
     4501 gggagatata tcatgaaagg ctggcttttt cttgttatcg caatagttgg cgaagtaatc
     4561 gcaacatccg cattaaaatc tagcgagggc tttactaagc tgatccggtg gatgaccttt
     4621 tgaatgacct ttaatagatt atattactaa ttaattgggg accctagagg tccccttttt
     4681 tattttaaaa attttttcac aaaacggttt acaagcatac tagaggatcg gcggccgcgg
     4741 agatctacgt acgcgtgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
     4801 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
     4861 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
     4921 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa
     4981 aaaacagact acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt
     5041 agatggtatt agtgacctgt agtcgaccga cagccttcca aatgttcttc gggtgatgct
     5101 gccaacttag tcgaccgaca gccttccaaa tgttcttctc aaacggaatc gtcgtatcca
     5161 gcctactcgc tattgtcctc aatgccgtat taaatcataa aaagaaataa gaaaaagagg
     5221 tgcgagcctc ttttttgtgt gacaaaataa aaacatctac ctattcatat acgctagtgt
     5281 catagtcctg aaaatcatct gcatcaagaa caacttcaca actcttatac ttttctctta
     5341 caagtcgttc ggcttcatct ggattttcag cctctatact tactaaacgt gataaagttt
     5401 ctgtaatttc tactgtatcg acctgcagac tggctgtgta taagggagcc tgacatttat
     5461 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca
     5521 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc
     5581 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc
     5641 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc
     5701 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
     5761 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac
     5821 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc
     5881 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac
     5941 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg
     6001 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat
     6061 acgcatactg ttatctggct tttagtaagc cggatccacg cggcgtttac gccccgccct
     6121 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     6181 agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     6241 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     6301 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     6361 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     6421 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     6481 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     6541 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     6601 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     6661 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     6721 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     6781 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     6841 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
     6901 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
     6961 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttagtc gactacaggt
     7021 cactaatacc atctaagtag ttgattcata gtgactggat atgttgtgtt ttacagtatt
     7081 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     7141 tttctcgttc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     7201 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     7261 tcccctatag tgagtcgtat tacatggtca tagctatttc ctggcagctc tggcccgtgt
     7321 ctcaaaatcc tcgaggccta ctagtcggat ccccgggtac cgagctcgaa tttcgagctt
     7381 ctggagcagg aagatgtcgc gggcattagc accagcggtc tgccaagcct ccgccagccg
     7441 ttgggtccct tccgcttgag cttttccatc ttcgacgata cgggcggcgg ccccccgcgc
     7501 ttccgcgatc gcccgtttac aagctgcctc agctggggcg atcacatcgg cttgaagttg
     7561 ctgctgcacc tgtttgatcc gctcctgctg cacagggagt tctgcttggc tacgagcgac
     7621 ttcggtagca atgtccgctt cagcttcggc caccaccgct tcgcgccgcg tcaacgcatc
     7681 ctgaatccgg cgctcggcct cggcttgggc gatcgctaca tcgcgatcga tccgacgcag
     7741 ggccgtgatc ttgtcatttt cggccgtttg gatcgcagag gcagcctggg catcggcttc
     7801 agcaattcgg gcatctcgct gcagatcagc ccgctgcttg cgtccactag ccgagagata
     7861 accgacctca tcggaaatgt tctggacttg cagcgtatcg aggactagac ccagctgctc
     7921 aaggtcatcc tccgcctctt ccagcagact tttggcaaag gcaattttgt cctcgttgat
     7981 ctgctccggc gtgaggctgg ctaaaacacc acgcaagttg ccttcgaggg tctccttggc
     8041 aatttgctcg atttccttac ggtttttgcc aagcagccgc tcgatcgcgt tgtggatggt
     8101 cggttcttcc ccagcaatct tgatattggc aacgccttca acagtcaggg gaatgccgcc
     8161 cttggagaag gcattggaaa cgcgcaactc aatgatcatg ttggtcagat ccatgcggag
     8221 cgctttttcc agcagaggta cccgcaggct gctgccgccc ttgaccaagc gatagccaac
     8281 tcggcggcca tcactactgc ggcgactact gccagcaaag atcaaaattt cactgggttg
     8341 gcagatgtag tagagattgc gcaggactaa gctgccagcc ccggcg
//
