LOCUS       Exported                5412 bp ds-DNA     circular SYN 09-JUN-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  AUTHORS   Karen Polizzi's Lab
  TITLE     Direct Submission
  JOURNAL   Exported Jun 9, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5412
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1230..1251
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     terminator      60..103
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
                     /note="color: #ffffff"
     promoter        297..327
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 297 .. 302 / #ffffff / -35
                        2: 303 .. 320 / #ffffff
                        3: 321 .. 327 / #ffffff / -10"
     RBS             369..380
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
                     /note="color: #a6acb3"
     CDS             387..1064
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="color: #ff0000"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     775..789
                     /label=SF_seq1
                     /note="color: black; sequence: cgtccgacggtccgg"
     terminator      1109..1180
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      1196..1223
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
                     /note="color: #ffffff"
     primer_bind     1310..1354
                     /label=SF_F1_fwd
                     /note="color: black; sequence: 
                     tatcagctcactcaaaggcggtaatTTAAGCCGAACAGGAAGCAC"
     primer_bind     complement(1315..1334)
                     /label=VR
                     /note="color: black; sequence: attaccgcctttgagtgagc"
     primer_bind     complement(1316..1334)
                     /label=EcoFlex_rev
                     /note="color: black; sequence: ATTACCGCCTTTGAGTGAG"
     rep_origin      complement(1428..2016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     1531..1547
                     /label=SF_seq2
                     /note="color: black; sequence: ggaagctccctcgtgcg"
     CDS             2220..3020
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /note="color: #ccffcc"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     primer_bind     2300..2316
                     /label=SF_seq3
                     /note="color: black; sequence: ggttacccccggcggtg"
     primer_bind     complement(2423..2440)
                     /label=SF_seq9
                     /note="color: black; sequence: cctgaccacgagggcatg"
     primer_bind     3081..3096
                     /label=SF_seq4
                     /note="color: black; sequence: gcaacacgtggagcgg"
     primer_bind     complement(3179..3196)
                     /label=SF_seq8
                     /note="color: black; sequence: ctaccgccggcgtaacag"
     oriT            3197..3306
                     /note="incP origin of transfer"
                     /note="color: #a6acb3"
     primer_bind     3789..3808
                     /label=SF_seq5
                     /note="color: black; sequence: gcttcacctgcataaatgag"
     CDS             complement(3882..5165)
                     /codon_start=1
                     /product="integrase"
                     /label=int
                     /note="color: #99cc00"
                     /translation="MAPRATNNPRQLRAKSCGCQLCMEKYPPEKYGERNRRRDCTGSWQ
                     ARYRDPAGNQKQKCFAIKDGGKKAAEAHLDKIRTQVRERTYADPKRGEITLSQWWKLWW
                     EAQPDRAVTTANRKRSNWAAHIEPKWGQWRLCDLEYIELQAWITKEVKGYHTRKKVHEV
                     LNSMLRAAVKDGRRIPFNPAADLDIGEAPAKHPDELMPPDRAQCALIVSHLPMYYRPLV
                     VFLEHTGLRWGEATALRWENVDLDAHHLKVKEVLSDDEGKLFRKPAPKSNAGFRTVPLT
                     PQAEDAIRTMVTRWRPTPTITPIGEDPYDLAPDELVFRGPQGGVLTRHNFRRTWIPAIK
                     AAGLAREVKNRDTGRMEWWPRVHDLRHVFATWLKDVGIDEKDTQTVMGHERGSKVTWLY
                     QHSPADVAAKVRAAMAPETEGVRTLRAV"
     primer_bind     complement(3894..3925)
                     /label=SF_F1_rev
                     /note="color: black; sequence: 
                     tggctcccgaaacCGAGGGTGTTCGAACGCTG"
     primer_bind     complement(3894..3912)
                     /label=SF_F0_rev
                     /note="color: black; sequence: 
                     atggtttcttagacgtcaggtggcaCGAGGGTGTTCGAACGCTG"
     primer_bind     3900..3932
                     /label=SF_F2_fwd
                     /note="color: black; sequence: 
                     tcgaacaccctcgGTTTCGGGAGCCATCGCCGC"
     primer_bind     complement(4458..4491)
                     /label=SF_F2_rev
                     /note="color: black; sequence: 
                     tgaacacaccggactcCGGTGGGGCGAGGCGACG"
     primer_bind     4463..4497
                     /label=SF_F3_fwd
                     /note="color: black; sequence: 
                     cctcgccccaccgGAGTCCGGTGTGTTCAAGgaag"
     primer_bind     4503..4522
                     /label=SF_seq6
                     /note="color: black; sequence: gagcggccggtagtacatcg"
     primer_bind     complement(4740..4773)
                     /label=SF_F3_rev
                     /note="color: black; sequence: 
                     gcagtggcgtctgTGCGACTTGGAGTACATCGAG"
     primer_bind     4748..4778
                     /label=SF_F4_fwd
                     /note="color: black; sequence: 
                     actccaagtcgcaCAGACGCCACTGCCCCCA"
     primer_bind     complement(join(5376..5412,1..12))
                     /label=SF_F4_rev
                     /note="color: black; sequence: 
                     atggtttcttagacgtcaggtggcaAGTACTCTGCTGCAACGTGTGAAC"
     primer_bind     complement(join(5400..5412,1..7))
                     /label=SF_seq7
                     /note="color: black; sequence: TTCTTAGACGTCAGGTGGCA"
     primer_bind     join(5400..5412,1..7)
                     /label=VF2
                     /note="color: black; sequence: tgccacctgacgtctaagaa"
     primer_bind     join(5400..5412,1..5)
                     /label=EcoFlex_fwd
                     /note="color: black; sequence: TGCCACCTGACGTCTAAG"
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa
       61 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct
      121 tctagaggtc tcactatatc tagagacgtc tagacatatg caatacgcaa accgcctctc
      181 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      241 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      301 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      361 tactagagaa agaggagaaa tactagatgg cttcctccga agacgttatc aaagagttca
      421 tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg
      481 aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg
      541 gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt
      601 acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca
      661 aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct
      721 ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg
      781 acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc
      841 cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc
      901 actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg
      961 cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg
     1021 aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaataacgc tgatagtgct
     1081 agtgtagatc gctactagag ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1141 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     1201 accttcgggt gggcctttct gcgtttatac gtctcattag gtacagagac cctgcaggct
     1261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1321 tcaaaggcgg taatttaagc cgaacaggaa gcacagctcc tactgaacat gtgagcaaaa
     1381 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1441 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1501 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1561 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1621 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1681 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1741 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1801 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1861 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1921 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1981 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2041 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2101 aaaaggatct tcacctagat ccttttggtt catgtgcagc tccatcagca aaaggggatg
     2161 ataagtttat caccaccgac tatttgcaac agtgccgttg atcgtgctat gatcgactga
     2221 tgtcatcagc ggtggagtgc aatgtcgtgc aatacgaatg gcgaaaagcc gagctcatcg
     2281 gtcagcttct caaccttggg gttacccccg gcggtgtgct gctggtccac agctccttcc
     2341 gtagcgtccg gcccctcgaa gatgggccac ttggactgat cgaggccctg cgtgctgcgc
     2401 tgggtccggg agggacgctc gtcatgccct cgtggtcagg tctggacgac gagccgttcg
     2461 atcctgccac gtcgcccgtt acaccggacc ttggagttgt ctctgacaca ttctggcgcc
     2521 tgccaaatgt aaagcgcagc gcccatccat ttgcctttgc ggcagcgggg ccacaggcag
     2581 agcagatcat ctctgatcca ttgcccctgc cacctcactc gcctgcaagc ccggtcgccc
     2641 gtgtccatga actcgatggg caggtacttc tcctcggcgt gggacacgat gccaacacga
     2701 cgctgcatct tgccgagttg atggcaaagg ttccctatgg ggtgccgaga cactgcacca
     2761 ttcttcagga tggcaagttg gtacgcgtcg attatctcga gaatgaccac tgctgtgagc
     2821 gctttgcctt ggcggacagg tggctcaagg agaagagcct tcagaaggaa ggtccagtcg
     2881 gtcatgcctt tgctcggttg atccgctccc gcgacattgt ggcgacagcc ctgggtcaac
     2941 tgggccgaga tccgttgatc ttcctgcatc cgccagaggc gggatgcgaa gaatgcgatg
     3001 ccgctcgcca gtcgattggc tgagctcatg agcggagaac gagatgacgt tggaggggca
     3061 aggtcgcgct gattgctggg gcaacacgtg gagcggatcg gggattgtct ttcttcagct
     3121 cgctgatgat atgccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct
     3181 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     3241 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     3301 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     3361 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     3421 cagggttatg cagcggaaaa gatccgtcga cctgcaggca tgctcctcta gagtccgtct
     3481 gacgcgtgtg gggagcacgc cggcccgctc ggtagacagc actctacgca ttgtggctgc
     3541 tgtgcacatc actctttcga gcgacgtgat ccgcaggatt ccgtggggat gtggcatcta
     3601 tgtggcatcc gggtacgaca acggcccccc gttcgaatgg aacggggggc cgttgactcc
     3661 ctgacctgcg atgcaggttg tgcccccggc aggattcgaa cctgcgacac ccgctttagg
     3721 agagaacgta aagcgatcat gagcctcagt gagtctgagt gagtctcagt gagcctgtcg
     3781 ccataggggc ttcacctgca taaatgagtt tcagtgagtc tcggtgagtc tcagtgaggg
     3841 ggtgttgtgg catccctgtg gcatctgcgt ggcatccggc gtcacaccgc ccgcagcgtt
     3901 cgaacaccct cggtttcggg agccatcgcc gcccgcacct tcgccgccac gtcggccggc
     3961 gaatgctggt acaaccacgt caccttcgac cctcgctcgt gacccatcac ggtctgcgtg
     4021 tccttctcgt caatgcccac atccttgagc cacgtggcga acacgtggcg aaggtcgtgc
     4081 acccgcggcc accactccat gcggccggtg tcccggttct tcacctcgcg ggcgaggcct
     4141 gcagccttga ttgcagggat ccatgtgcgc cggaagttgt gccgggtcag gacgccgccc
     4201 tgtgggccgc ggaacacgag ctcatccggc gcgaggtcgt acgggtcctc gccaatcggg
     4261 gtgatcgtgg gagtcggccg ccaccgggtg accatggtgc ggatcgcgtc ctcggcctgc
     4321 ggcgtgagcg ggaccgtgcg gaacccggcg ttgctcttcg gcgcaggctt ccggaacagc
     4381 ttgccttcgt cgtcactgag cacttccttc accttgaggt ggtgggcgtc caggtcgacg
     4441 ttctcccagc gcagcgccgt cgcctcgccc caccggagtc cggtgtgttc aaggaagacg
     4501 acgagcggcc ggtagtacat cggcaggtga ctgacgatca gcgcgcactg cgcgcggtcg
     4561 ggcggcatca gttcgtccgg atgcttcgcc ggcgcctcgc caatgtccag gtcggccgcc
     4621 gggttgaacg ggatacgccg gccgtccttg acggcggccc ggagcatcga gttcagcacc
     4681 tcatgaacct tcttccgggt gtggtagccc ttcacctcct tcgtgatcca cgcctgcagc
     4741 tcgatgtact ccaagtcgca cagacgccac tgcccccact tcggctcgat gtgcgcggcc
     4801 cagttcgacc gcttccggtt ggcggtcgtg actgctcggt ccggctgcgc ctcccaccac
     4861 agtttccacc actgggacag ggtgatctcg ccacgcttcg ggtcggcgta cgtccgttcg
     4921 cggacctgcg tgcggatctt gtcgaggtgc gcctcggctg ccttcttacc gccgtccttg
     4981 atcgcaaagc atttctgctt ctggttgccg gccgggtcgc ggtaacgcgc ctgccacgag
     5041 ccggtgcagt cgcgtcggcg gttccgctct ccgtacttct cgggcgggta cttctccatg
     5101 cacagctggc agccgcagct cttcgccctc aactgccgtg ggttgttcgt cgctctaggc
     5161 gccatgggtc acctgtccgc tcctccgctg ttcgggaacg cggggcagga tgtgtacggc
     5221 tgccccacac cagcagactg caccgtccgg gggctgctct acagccagtt cttcaaggac
     5281 ggctcgaagg gctacgaggg aagcatcccg ggggagggtg gcaggcaggg tgaccgtgtt
     5341 tccagcggcg tcgtacggcc ggatgacact cgttcgttca cacgttgcag cagagtactt
     5401 gccacctgac gt
//
