LOCUS       Exported                4723 bp ds-DNA     circular SYN 20-OCT-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4723)
  AUTHORS   EMBL Monterotondo
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4723
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(52..69)
                     /label=MP1.1
                     /note="color: black; sequence: gggcgcgtcccattcgcc; added: 
                     2020-09-24"
     rep_origin      62..69
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     70..94
                     /label=MP1.2
                     /note="color: black; sequence: gagttttcgttccactgagcgtcag;
                     added: 2020-09-23"
     rep_origin      123..711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(725..747)
                     /label=2.77
                     /note="color: black; sequence: 
                     caattcgttcaagccgagatcggggccaggaaccgtaaaaaggccg; added: 
                     2023-10-20"
     primer_bind     748..774
                     /label=2.70
                     /note="color: black; sequence: ttttgctggccttttgctcacatgttc;
                     added: 2023-10-20"
     primer_bind     748..772
                     /label=2.18
                     /note="color: black; sequence: 
                     ttttgctggccttttgctcacatgtATAACTTCGTATAGGATACTTTATACGAAGTTAT
                      cgctcactgactcgctgc; added: 2023-10-20"
     primer_bind     complement(752..772)
                     /label=1.96
                     /note="color: black; sequence: acatgtgagcaaaaggccagc;
                     added: 2023-10-20"
     primer_bind     complement(863..881)
                     /label=2.18
                     /note="color: black; sequence: 
                     ttttgctggccttttgctcacatgtATAACTTCGTATAGGATACTTTATACGAAGTTAT
                      cgctcactgactcgctgc; added: 2023-10-20"
     mobile_element  1016..1240
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
                     /note="color: #ffe4c4"
     primer_bind     complement(1284..1306)
                     /label=2.77
                     /note="color: black; sequence: 
                     caattcgttcaagccgagatcggggccaggaaccgtaaaaaggccg; added: 
                     2023-10-20"
     primer_bind     1284..1306
                     /label=2.76
                     /note="color: black; sequence: ccgatctcggcttgaacgaattg;
                     added: 2023-10-20"
     CDS             complement(1307..1840)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /note="color: #ccffcc"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(2029..2057)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
                     /note="This reverse directional feature has 3 segments:
                     1: 2029 .. 2034 / #ffffff / -10
                     2: 2035 .. 2051 / #ffffff
                     3: 2052 .. 2057 / #ffffff / -35"
     primer_bind     2050..2064
                     /label=2.74
                     /note="color: black; sequence: 
                     tatgtcaactgggttcgtgccttcgccgcggtaccataacttcgtatagc; added: 
                     2023-10-20"
     primer_bind     complement(2213..2233)
                     /label=1.65
                     /note="color: black; sequence: 
                     gatctattaatattccggagtaggtcgATCCGTGCACAGCACCTTGC; added: 
                     2023-10-20"
     primer_bind     2428..2452
                     /label=2.16
                     /note="color: black; sequence: agatcctctacgcggccgcggtacc;
                     added: 2023-10-20"
     primer_bind     complement(2435..2452)
                     /label=2.14
                     /note="color: black; sequence: ggtaccgcggccgcgtag; added: 
                     2023-10-20"
     primer_bind     2442..2467
                     /label=2.74
                     /note="color: black; sequence: 
                     tatgtcaactgggttcgtgccttcgccgcggtaccataacttcgtatagc; added: 
                     2023-10-20"
     protein_bind    2453..2486
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     primer_bind     complement(2460..2492)
                     /label=2.9
                     /note="color: black; sequence: 
                     aaccagataacttcgtataatgtatgctatacg; added: 2023-10-20"
     primer_bind     2493..2520
                     /label=2.11
                     /note="color: black; sequence: 
                     gttcggtcccagggctggcactctgttactagtatcgattcgcgacctactcc; 
                     added: 2023-10-20"
     misc_feature    2510..2536
                     /label=5' annealing
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     complement(2510..2536)
                     /label=MP1.3
                     /note="color: black; sequence: gatctattaatattccggagtaggtcg;
                     added: 2020-09-24"
     primer_bind     2510..2536
                     /label=CasI_for
                     /note="color: black; sequence: 
                     AACGCTCTATGGTCTAAAGATTTAAATCGACCTACTCCGGAATATTAATAGATC; 
                     added: 2021-01-06"
     primer_bind     2510..2536
                     /label=CasII_for
                     /note="color: black; sequence: 
                     AAACTGGATACTATTGCACGTTTAAATCGACCTACTCCGGAATATTAATAGATC; 
                     added: 2021-01-06"
     primer_bind     2510..2536
                     /label=CasIII_for
                     /note="color: black; sequence: 
                     AAACCTAATGATGCCTGATGTTTAAATCGACCTACTCCGGAATATTAATAGATC; 
                     added: 2021-01-06"
     primer_bind     2510..2536
                     /label=CasIV_for
                     /note="color: black; sequence: 
                     AAACGGTTCACATAGCTTAGTTTAAATCGACCTACTCCGGAATATTAATAGATC; 
                     added: 2021-01-06"
     primer_bind     2510..2536
                     /label=CasV_for
                     /note="color: black; sequence: 
                     AAACACTGACATTGACTTGGTTTAAATCGACCTACTCCGGAATATTAATAGATC; 
                     added: 2021-01-06"
     primer_bind     complement(2510..2536)
                     /label=1.65
                     /note="color: black; sequence: 
                     gatctattaatattccggagtaggtcgATCCGTGCACAGCACCTTGC; added: 
                     2023-10-20"
     primer_bind     2512..2557
                     /label=3.67
                     /note="color: black; sequence: 
                     acctactccggaatattaatagatcACTCGAGGGATCCAAGCTggC; added: 
                     2023-09-14"
     primer_bind     2537..2556
                     /label=2.96
                     /note="color: black; sequence: 
                     GAAAGATAGTCTGCGTAAAATTGACGCATGACTCGAGGGATCCAAGCTgg; added: 
                     2023-10-20"
     promoter        2566..2957
                     /label=Second generation TRE promoter
                     /note="color: #ffffff"
     protein_bind    2566..2584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2603..2621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2640..2658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2677..2695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2751..2769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2788..2806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    2825..2843
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     primer_bind     complement(2956..2999)
                     /label=3.68
                     /note="color: black; sequence: 
                     ccgcgcccgatggtgggacggtatgACTTGTGATGGCCGCCACC; added: 
                     2023-09-14"
     primer_bind     2975..2994
                     /label=3.66
                     /note="color: black; sequence: cataccgtcccaccatcggg; added:
                     2023-09-14"
     primer_bind     2976..2995
                     /label=1.15
                     /note="color: black; sequence: ataccgtcccaccatcgggc; added:
                     2023-10-20"
     primer_bind     3009..3051
                     /label=3.79
                     /note="color: black; sequence: 
                     cgaagcgcgcggaattcaaaggATGAGGCTTCGTGAACCCTTG; added: 
                     2023-10-20"
     primer_bind     complement(3013..3030)
                     /label=1.34
                     /note="color: black; sequence: CCTTTGAATTCCGCGCGC; added: 
                     2023-10-20"
     CDS             3298..3993
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /note="color: #3366ff"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     complement(3978..4025)
                     /label=3.80
                     /note="color: black; sequence: 
                     actagtgagctcgtcgacgtaggCTAGGATCCATTAAGCTTGTGCCCC; added: 
                     2023-10-20"
     primer_bind     4003..4024
                     /label=2.19
                     /note="color: black; sequence: cctacgtcgacgagctcactag;
                     added: 2023-10-20"
     primer_bind     complement(4039..4062)
                     /label=2.27
                     /note="color: black; sequence: GTCGAGACTGCAGGCTCTAGATTC;
                     added: 2023-10-20"
     primer_bind     4042..4062
                     /label=2.26
                     /note="color: black; sequence: TCTAGAGCCTGCAGTCTCGAC;
                     added: 2023-10-20"
     primer_bind     4144..4165
                     /label=MP1.4
                     /note="color: black; sequence: cacctccccctgaacctgaaac;
                     added: 2020-09-24"
     primer_bind     complement(4144..4165)
                     /label=3.7
                     /note="color: black; sequence: GTTTCAGGTTCAGGGGGAGGTG;
                     added: 2023-10-20"
     polyA_signal    4192..4326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     4399..4448
                     /label=1.60
                     /note="color: black; sequence: 
                     cgctgagcaataactatcataaccCCTAGGAGATCCGAACCAGATAAGTG; added: 
                     2023-10-20"
     misc_feature    complement(4399..4422)
                     /label=3' annealing
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     complement(4399..4422)
                     /label=Cas?_rev
                     /note="color: black; sequence: 
                     AACCCCGATTGAGATATAGATTTATTTAAATGGTTATGATAGTTATTGCTCAGCG; 
                     added: 2021-01-06"
     primer_bind     complement(4399..4422)
                     /label=CasI_rev
                     /note="color: black; sequence: 
                     AAACGTGCAATAGTATCCAGTTTATTTAAATGGTTATGATAGTTATTGCTCAGCG; 
                     added: 2021-01-06"
     primer_bind     complement(4399..4422)
                     /label=CasII_rev
                     /note="color: black; sequence: 
                     AAACATCAGGCATCATTAGGTTTATTTAAATGGTTATGATAGTTATTGCTCAGCG; 
                     added: 2021-01-06"
     primer_bind     complement(4399..4422)
                     /label=CasIII_rev
                     /note="color: black; sequence: 
                     AAACTAAGCTATGTGAACCGTTTATTTAAATGGTTATGATAGTTATTGCTCAGCG; 
                     added: 2021-01-06"
     primer_bind     complement(4399..4422)
                     /label=CasIV_rev
                     /note="color: black; sequence: 
                     AAACCAAGTCAATGTCAGTGTTTATTTAAATGGTTATGATAGTTATTGCTCAGCG; 
                     added: 2021-01-06"
     primer_bind     4423..4448
                     /label=2.8
                     /note="color: black; sequence: cctaggagatccgaaccagataagtg; 
                     added: 2023-10-20"
     mobile_element  4436..4601
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
                     /note="color: #ffe4c4"
     primer_bind     4580..4602
                     /label=2.15
                     /note="color: black; sequence: ccaactattttgtccgcccacag;
                     added: 2023-10-20"
     primer_bind     complement(4580..4602)
                     /label=2.17
                     /note="color: black; sequence: ctgtgggcggacaaaatagttgg;
                     added: 2023-10-20"
ORIGIN
        1 ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa tggcgaatgg
       61 gacgcgcccg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      241 ttcagcagag cgcagatacc aaatactatt cttctagtgt agccgtagtt aggccaccac
      301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      781 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      841 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
      901 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt
      961 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg
     1021 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     1081 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     1141 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     1201 aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac
     1261 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     1321 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     1381 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     1441 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     1501 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     1561 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     1621 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     1681 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     1741 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     1801 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     1861 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     1921 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     1981 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     2041 gaacaggctt atgtcaactg ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     2101 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     2161 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     2221 ctgtgcacgg atctgccctt gcttcaggag atcggtagac ctcggccgtc gcggcgcttg
     2281 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     2341 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacagct ttgtttgtac
     2401 tatcaacagg ttgaactgct gatcaacaga tcctctacgc ggccgcggta ccataacttc
     2461 gtatagcata cattatacga agttatctgg ttttactagt atcgattcgc gacctactcc
     2521 ggaatattaa tagatcactc gagggatcca agctggccgc acgtctccct atcagtgata
     2581 gagaagtcga cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct
     2641 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     2701 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     2761 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     2821 gagctcccta tcagtgatag agaaggtacc ccctatataa gcagagctcg tttagtgaac
     2881 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     2941 cgatccagcc tccgcggtgg cggccatcac aagtcatacc gtcccaccat cgggcgcgga
     3001 tcccggtccg aagcgcgcgg aattcaaagg atgaggcttc gtgaaccctt gttgtcaggc
     3061 tctgctgcca tgcccggtgc cagcctgcaa agggcatgtc gtcttctggt tgcggtgtgt
     3121 gcattgcatc tcggggtgac tcttgtgtat tatcttgcgg gaagagatct ctcccggctt
     3181 ccacagttgg tgggcgtgag tacccccttg caaggcggtt caaatagcgc tgcagctatt
     3241 ggacaaagca gcggcgaact gaggacaggc ggcgctaaag acccaccagt agcgactagc
     3301 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat
     3361 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     3421 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc
     3481 ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag
     3541 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     3601 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     3661 ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     3721 ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg
     3781 aagctcgtgg gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa
     3841 cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc
     3901 aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc
     3961 gacctcccta gcaaactggg gcacaagctt aatggatcct agcctacgtc gacgagctca
     4021 ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt cgagaagtac
     4081 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4141 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4201 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4261 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4321 tggatctgat cactgcttga gcctagaaga tccggctgct aacaaagccc gaaaggaagc
     4381 tgagttggct gctgccaccg ctgagcaata actatcataa cccctaggag atccgaacca
     4441 gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat tagcttacga
     4501 cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt aaaaactcca
     4561 tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat ttttcttcct
     4621 gttatgtttt taatcaaaca tcctgccaac tccatgtgac aaaccgtcat cttcggctac
     4681 tttttctctg tcacagaatg aaaatttttc tgtcatctct tcg
//
