LOCUS       Exported                5418 bp DNA     circular SYN 23-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5418)
  AUTHORS   EJPARKER
  TITLE     Direct Submission
  JOURNAL   Exported Apr 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   274..302
                     /label=IR1
     rep_origin      587..975
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     complement(1350..1377)
                     /label=pHimarEm1+XbaI_RV2
     primer_bind     1357..1399
                     /label=Fragment.FOR
     primer_bind     1380..1405
                     /locus_tag=""
                     /label=Geneious name: MCS primer
                     /note="Geneious name: MCS primer"
     primer_bind     1380..1405
                     /label=pProbeMCSfor
     primer_bind     1380..1405
                     /label=pFru97 MCS fw
     primer_bind     1380..1403
                     /label=cassette_pAG408overhang_FWD
     primer_bind     1380..1400
                     /label=REV_pFru_ins
     primer_bind     1380..1399
                     /label=cassette5_pGRG36overhang_FWD
     primer_bind     1390..1426
                     /label=FWD_primer_PnptII
     promoter        1417..1753
                     /locus_tag=""
                     /label=p-nptII promoter
                     /label=p-nptII
     primer_bind     1417..1436
                     /label=11x-nptII-FL-fw
     primer_bind     1417..1431
                     /label=lederm_nptII.fw
     primer_bind     complement(1566..1590)
                     /label=pntpII_1_REV
     primer_bind     1566..1590
                     /label=pntpII_1_FWD
     primer_bind     1567..1585
                     /label=pFru97xpMRE100_nptIIGib_rv
     primer_bind     complement(1688..1708)
                     /label=pntpII_2_REV
     primer_bind     1688..1708
                     /label=pntpII_2_FWD
     primer_bind     complement(1732..1766)
                     /label=rbs_1_REV
     primer_bind     1732..1766
                     /label=rbs_1_FWD
     primer_bind     complement(1752..1784)
                     /label=REV_primer_PnptII
     primer_bind     complement(1752..1781)
                     /label=REV_primer_PnptII (3)
     primer_bind     complement(1752..1777)
                     /label=REV_primer_PnptII (2)
     primer_bind     1770..1787
                     /label=XhoI_Ptac_XFP_for
     CDS             1772..2467
                     /codon_start=1
                     /locus_tag=""
                     /product=""
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     1772..1789
                     /label=XFPs for
     primer_bind     complement(1772..1789)
                     /label=Ptuf rev
     primer_bind     complement(2448..2487)
                     /label=REV_primer_mTq2
     primer_bind     complement(2449..2471)
                     /label=NotI_XFP_rev
     primer_bind     complement(2449..2470)
                     /label=nextgenFP.rv
     primer_bind     complement(2451..2487)
                     /label=REV_primer_mO
     primer_bind     complement(2451..2470)
                     /label=11x-FL-rv
     primer_bind     complement(2452..2487)
                     /label=REV_primer_sY2
     primer_bind     complement(2452..2470)
                     /label=REV_sYFP2
     primer_bind     complement(2452..2470)
                     /label=Ptac+eXFP_rev
     primer_bind     complement(2453..2470)
                     /label=XFPs rev KpnI
     primer_bind     2468..2482
                     /label=Gib_Gent.rev
     primer_bind     complement(2475..2494)
                     /label=FWD_pFru_ins
     primer_bind     2478..2500
                     /label=Gib_Gent.for.cor
     terminator      2484..2578
                     /locus_tag=""
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2622..3281
                     /codon_start=1
                     /locus_tag=""
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2683..2711)
                     /label=pMRE130inserts_rev
     primer_bind     complement(3266..3281)
                     /label=Chloramp_rev
     primer_bind     3316..3336
                     /label=ARB-RB-PCR1
     terminator      3346..3432
                     /locus_tag=""
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB ge
                     ne"
     primer_bind     3418..3438
                     /label=ARB-RB-PCR2
     primer_bind     complement(3437..3454)
                     /label=Gib_Gent.for
     primer_bind     complement(3442..3481)
                     /label=Fragment.REV
     primer_bind     complement(3442..3459)
                     /label=cassette_pGRG36overhang_RV
     primer_bind     complement(3442..3459)
                     /label=pMRE130_cassette2_RV
     primer_bind     complement(3443..3459)
                     /label=cassette_pAG408overhang_RV
     primer_bind     3460..3486
                     /label=pHimarEm1+XmaI_FWD
     repeat_region   3549..3577
                     /label=IR2
     primer_bind     3823..3839
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3919..3930)
                     /label=cassette_pAG408overhang_RV
     oriT            3926..4034
                     /note="incP origin of transfer"
     protein_bind    4181..4202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4217..4247
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        4217..4246
                     /gene="lac prom"
                     /label=lac prom promoter
     protein_bind    4255..4271
                     /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)."
     misc_feature    4291..5337
                     /label=Transposase
                     /label=nonstandard type:  ORF
                     /note="/sequence=ORF_2 rf(1)"
ORIGIN
        1 ctagatgcat gctcgagcgg ccgccagtgt gatggatttg aaaaaggaag agtatgagta
       61 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      241 gttttccaat gatgagcact tttaaagttc tgctaacagg ttggctgata agtccccggt
      301 ctgacacata gatggcgtcg ctagtattaa atgcatatta tttttatata gtaccaacct
      361 tcaaatgatt cccagatctc aaactggaac aacactcaac cctatctcgg tctattcttt
      421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      481 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
      541 ggaaatgtgc gcggaaccag atccccacga tgcgtccggc gtagaggatc tgaagatcag
      601 cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta agctctcatg
      661 tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac gtactaagct
      721 ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg tttgaacaat
      781 aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt ataagaaaaa
      841 aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca acaagccagg
      901 gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttcagtg acacaggaac
      961 acttaacggc tgacagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     1021 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggttaatt
     1081 aagggctgca ggaattcgat atcaagctta tcgataccgt cgaaattctg attaataatt
     1141 tgtttaaatt tttcgtttgg cgtgaggtat ccaagtcttt tacgaggtcg attattgagt
     1201 ttattttcaa tccacttaat ctgtttgttg gttacttcac taaagtcctt accctttggg
     1261 atatactgcc tgataagccc gttggtgttt tcattggcac cacgttccca tgagtggtat
     1321 ggtttgcaaa aatagaattt tatttccaat ttttgcgcaa tttcctcgtg ctctagaata
     1381 taaactgcca ggaattgggg atcggattcg agctcgcacg ctgccgcaag cactcagggc
     1441 gcaagggctg ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac
     1501 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
     1561 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtc ttatggacag
     1621 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
     1681 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc
     1741 aagagacagg atactagtgg aggaagaaaa aatggtgagc aagggcgagg cagtgatcaa
     1801 ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat
     1861 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1921 caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc
     1981 cagggccttc atcaagcacc ccgccgacat ccccgactac tataagcagt ccttccccga
     2041 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca
     2101 ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt
     2161 ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg
     2221 gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga
     2281 cggcggccgc tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat
     2341 gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc tcccacaacg aggactacac
     2401 cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct
     2461 gtacaagtaa ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact
     2521 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     2581 aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     2641 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     2701 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     2761 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     2821 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     2881 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     2941 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     3001 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     3061 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     3121 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     3181 tggcgattca ggttcatcat gccgtctgtg atggcttcca tgtcggcaga atgcttaatg
     3241 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
     3301 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
     3361 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     3421 agtaggacaa atccgccgct aggagcttgc ggccgccagc ccgggcaatt cgaggggtat
     3481 cgctcttgaa gggaactatg ttgaataata aaaacgaatt ttgacaaaaa aatgtgtttt
     3541 tctttgttag accggggact tatcagccaa cctgttatgt ggcgcggtat tatcccgtat
     3601 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3661 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3721 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcgaggc
     3781 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     3841 aattaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3901 gataataatg gtttcttaga gcttacggcc agcctcgcag agcaggattc ccgttgagca
     3961 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     4021 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     4081 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     4141 ccccgaagca gggttatgca gcgggtgagc gcaacgcaat taatgtgagt tagctcactc
     4201 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     4261 gcggataaca atttcacaca ggaaacacat atggaaaaaa aggaatttcg tgttttgata
     4321 aaatactgtt ttctgaaggg aaaaaatacg gtggaagcaa aaacttggct tgataatgag
     4381 tttccggact ctgccccagg gaaatcaaca ataattgatt ggtatgcaaa attcaagcgt
     4441 ggtgaaatga gcacggagga cggtgaacgc agtggacgcc cgaaagaggt ggttaccgac
     4501 gaaaacatca aaaaaatcca caaaatgatt ttgaatgacc gtaaaatgaa gttgatcgag
     4561 atagcagagg ccttaaagat atcaaaggaa cgtgttggtc atatcattca tcaatatttg
     4621 gatatgcgga agctctgtgc aaaatgggtg ccgcgcgagc tcacatttga ccaaaaacaa
     4681 caacgtgttg atgattctga gcggtgtttg cagctgttaa ctcgtaatac acccgagttt
     4741 ttccgtcgat atgtgacaat ggatgaaaca tggctccatc actacactcc tgagtccaat
     4801 cgacagtcgg ctgagtggac agcgaccggt gaaccgtctc cgaagcgtgg aaagactcaa
     4861 aagtccgctg gcaaagtaat ggcctctgtt ttttgggatg cgcatggaat aatttttatc
     4921 gattatcttg agaagggaaa aaccatcaac agtgactatt atatggcgtt attggagcgt
     4981 ttgaaggtcg aaatcgcggc aaaacggccc cacatgaaga agaaaaaagt gttgttccac
     5041 caagacaacg caccgtgcca caagtcattg agaacgatgg caaaaattca tgaattgggc
     5101 ttcgaattgc ttccccaccc accgtattct ccagatctgg cccccagcga ctttttcttg
     5161 ttctcagacc tcaaaaggat gctcgcaggg aaaaaatttg gctgcaatga agaggtgatc
     5221 gccgaaactg aggcctattt tgaggcaaaa ccgaaggagt actaccaaaa tggtatcaaa
     5281 aaattggaag gtcgttataa tcgttgtatc gctcttgaag ggaactatgt tgaataataa
     5341 aaacgaattt tgacaaaaaa atgtgttttt ctttgttaga ccggaatctg cagaattcca
     5401 gcacactggc ggccgtta
//
