LOCUS       62056_17165        9544 bp DNA     circular SYN 05-AUG-2020
DEFINITION  Cloning vector pMM7-10-1, complete sequence.
ACCESSION   MN630871
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9544)
  AUTHORS   Das SR, Maselko MB, Upadhyay A, Smanski MJ.
  TITLE     Batch cultivation of non-transgenic males suitable for juvenile 
            release SIT programs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9544)
  AUTHORS   Das SR, Maselko MB, Upadhyay A, Smanski MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2019) Biotechnology Institute, University of 
            Minnesota, 1479 Gortner Ave, St Paul, MN 55108, USA
REFERENCE   3  (bases 1 to 9544)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-OCT-2019) Biotechnology Institute, University of Minnesota, 1479
            Gortner Ave, St Paul, MN 55108, USA"
FEATURES             Location/Qualifiers
     source          1..9544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(24..42)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(49..65)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(206..661)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        688..792
                     /label=AmpR promoter
     CDS             793..1650
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1824..2412
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2700..2721
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2736..2766
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2774..2790
                     /label=lac operator
                     /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)."
     primer_bind     2798..2814
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2835..2853
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(3072..3141)
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     protein_bind    complement(3685..3963)
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     regulatory      3983..4135
                     /label=Hsp70
                     /note="Hsp70"
                     /regulatory_class="promoter"
     intron          4146..4212
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     5'UTR           4234..4254
                     /label=syn21 5'UTR
                     /note="syn21 5'UTR"
     CDS             4255..4878
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             5035..5268
                     /label=VP16 AD
                     /note="transcriptional activation domain of herpes simplex
                     virus protein VP16 (Triezenberg et al., 1988; Cousens et 
                     al., 1989)"
     intron          5356..5421
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5551..5571
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    5996..6128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        6396..8204
                     /label=Polyubiquitin
                     /note="Promoter of polyubiqutin gene of Drosophila
                     melanogaster"
     CDS             8229..8249
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             8433..9149
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    9153..9393
                     /label=terminator
                     /note="terminator"
     promoter        complement(9462..9480)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9511..9529
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcccggtac gtacccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
       61 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      121 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      181 gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg
      241 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
      301 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
      361 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
      421 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
      481 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
      541 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
      601 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
      661 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      721 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      781 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      841 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      901 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      961 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1021 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1081 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1141 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1201 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1261 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1321 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1381 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1441 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1501 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1561 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1621 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1681 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1741 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1801 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1921 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1981 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2041 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2101 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2161 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2221 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2281 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2341 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2401 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2461 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2521 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2581 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2641 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2701 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2761 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     2821 tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggctag aactagtgtc
     2881 gacatgcccg ccgtgaccgt cgagaacccg ctgacgctgc cccgcgtatc cgcacccgcc
     2941 gacgccgtcg cacgtcccgt gctcaccgtg accaccgcgc ccagcggttt cgagggcgag
     3001 ggcttcccgg tgcgccgcgc gttcgccggg atcaactacc gccacctcga cccgttcatc
     3061 atgatggacc agatgggtga ggtggagtac gcgcccgggg agcccaaggg cacgccctgg
     3121 cacccgcacc gcggcttcga gaccgtgacc tacatcgtcg acactagtgg atccagcggc
     3181 cgcacctgca ggccggccgt taacacgcgt ccgcggtcta gactcgagga ttccagatct
     3241 ggtaccgggc cgctgtatgg atatttgcag ggccgcagaa ttctctctat cactgatagg
     3301 gaggtctcta tcactgatag ggagttctct atcactgata gggatgtctc tatcactgat
     3361 agggatttct ctatcactga tagggaagtc tctatcactg atagggacct ctctatcact
     3421 gatagggaaa tctctatcac tgatagggat ctctctatca ctgataggga cttctctatc
     3481 actgataggg acgtctctat cactgatagg gaactctcta tcactgatag ggacatctct
     3541 atcactgata gggacttctc tatcactgat agggaagtat gttctctctc ttctcttctc
     3601 tctctctttc tcgaatgttc tctctcttct cttctctctc tctttctcga tggccggggc
     3661 gcgccaggtt tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt
     3721 cacttttctc tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg
     3781 atagggagtg gtaaactcga ctttcacttt tctctatcac tgatagggag atccgagctc
     3841 gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc gactttcact
     3901 tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta tcactgatag
     3961 ggagtggtaa actcgaagcg ccggatccgt cgagggaaaa gagcgccgga gtataaatag
     4021 aggcgcttcg tctacggagc gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa
     4081 gcgaaagcta agcaaataaa caagcgcagc tgaacaagct aaacaatctg cagccgatct
     4141 aaaaggtagg ttcaaccact gatgcctagg cacaccgaaa cgactaaccc taattcttat
     4201 cctttacttc aggcggccgg gctcgagggt accaacttaa aaaaaaaaat caaaatggtc
     4261 agccgtttgg ataaatccaa agttattaat tccgctttgg aattgttgaa tgaagttggt
     4321 attgaaggtt tgacaacacg taaattggct caaaaattgg gtgttgaaca accaacattg
     4381 tattggcatg ttaaaaataa acgtgctttg ttggatgctt tggctattga aatgttggac
     4441 cgtcatcata cacatttttg cccattggaa ggcgaatcct ggcaagattt cttgcgtaat
     4501 aatgccaaat ccttccgttg tgctttgttg tcccatcgtg atggtgccaa ggttcatttg
     4561 ggcacacgtc caacagaaaa acaatatgaa acattggaaa atcaattggc tttcttgtgt
     4621 caacaaggct tcagcttgga aaatgctttg tatgctttga gcgccgttgg tcattttaca
     4681 ttgggctgtg tgttggaaga tcaagaacat caagtcgcta aagaagaacg tgaaacacca
     4741 acaacagatt cgatgccccc attgttgcgt caagcaattg aattgttcga tcatcaagga
     4801 gccgaaccag cattcttgtt cggcttggaa ttgattattt gtggattgga aaaacaattg
     4861 aaatgtgaat cgggctcggg ccccgcgtac agccgcgcgc gtacgaaaaa caattacggg
     4921 tctaccatcg agggcctgct cgatctcccg gacgacgacg cccccgaaga ggcggggctg
     4981 gcggctccgc gcctgtcctt tctccccgcg ggacacacgc gcagactgtc gacggccccc
     5041 ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg
     5101 catgccgacg cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt
     5161 ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag
     5221 tttgagcaga tgtttaccga tgcccttgga attgacgagt acggtgggta gtaagcttgg
     5281 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
     5341 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt
     5401 ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg
     5461 aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag
     5521 gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc
     5581 aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact
     5641 cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt
     5701 atggaaaaat attctgtaac ctttataagt aggcataaca gttataatca taacatactg
     5761 ttttttctta ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg
     5821 tgtaccttta gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc
     5881 ttgactagag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5941 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     6001 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6061 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6121 tgtctcgagc atgcgcaaat ttaaagcgct gatatcgatc gcgcgcagat ctgtcatgat
     6181 gatcattgca attctgcagt cgacggtacc cgatcttgtc gccggaacgc agcgacagag
     6241 attccaatgt gtccgtatct ttcaggcttt tgcccttcag ttccagacga agcgactggc
     6301 gattcgcgtg tggggtctgc ttcagggtct tgtgaattag ggcgcgcaga tcgccgatgg
     6361 gcgtggcgcc ggagggcacc ttcaccttgc cgtacggctt gctgttcttc gcgttcaaaa
     6421 tctccagctc cattttgctt tcggtgcgct tgcaatcagt actgtccaaa atcgaaaatc
     6481 gccgaaccgt agtgtgaccg tgcggggctc tgcgaaaata aactttttta ggtatatggc
     6541 cacacacggg gaaagcacag tggattatat gttttaatat tataatatgc aggttttcat
     6601 tacttatcca gatgtaagcc cacttaaagc gatttaacaa ttatttgccg aaagagtaaa
     6661 aacaaatttc acttaaaaat ggattaagaa aagcttgtgt aagattatgc gcagcgttgc
     6721 cagatagctc catttaaaac acttcaaaaa caataagttt tgaaaatata tacataaata
     6781 gcagtcgttg ccgcaacgct caacacatca cacttttaaa acacccttta cctacacaga
     6841 attacttttt aaatttccag tcaagctgcg agtttcaaaa ttatagccgg tagagaagac
     6901 agtgctattt caaaagcaaa ctaaataaac accaatccta acaagccttg gacttttgta
     6961 agtttagatc aaaggtggca ttgcattcaa tgtcatggta agaagtaggt cgtctaggta
     7021 gaaatcctca ttcagccggt caagtcagta cgagaaaggt ctcaatttga aattgtctta
     7081 aaaatatttt attgttttgt actgtggtga gtttaaacga aaaacacaaa aaaaaagtga
     7141 tacacagaaa tcataaaaaa ttttaataca aggtattcgt acgtatcaaa aacatttcgg
     7201 cacaattttt tttctctgta ctaaagtgtt acgaacacta cggtattttt tagtgatttt
     7261 caacggacac cgaaggtata taaacagcgt tcgcgaacgg tcgccttcaa aaccaattga
     7321 catttgcagc agcaagtaca agcagaaagt aaagcgcaat cagcgaaaaa tttatactta
     7381 attgttggtg attaaagtac aattaaaaga acattctcga aagtcacaag aaacgtaagt
     7441 ttttaactcg ctgttaccaa ttagtaataa gagcaacaag acgttgagta atttcaagaa
     7501 aaactgcatt tcaaggtctt tgttcggcca tttttttttt attcaacgct ctacgtaatt
     7561 acaaaataag aaattggcag ccacgcatct tgttttccca atcaattggc atcaaaacgc
     7621 aaacaaatct ataaataaaa cttgcgtgtt gattttcgcc aagatttatt ggcaaattgt
     7681 gaaattcgca gtgacgcatt tgaaaattcg agaaatcacg aacgcactcg agcatttgtg
     7741 tgcatgttat tagttagtta gttctttgct taattgaagt attttaccaa cgaaatccac
     7801 ttatttttag ctgaaataga gtaggttgct tgaaacgaaa gccacgtctg gaaaatttct
     7861 tattgcttag tagttgtgac gtcaccatat acacacaaaa taatgtgtat gcatgcgttt
     7921 cagctgtgta tatatacatg cacacactcg cattatgaaa acgatgacga gcaacggaac
     7981 aggtttctca actacctttg ttcctgtttc ttcgctttcc tttgttccaa tattcgtaga
     8041 gggttaatag gggtttctca acaaagttgg cgtcgataaa taagtttccc atttttattc
     8101 cccagccagg aagttagttt caatagtttt gtaatttcaa cgaaactcat ttgatttcgt
     8161 actaattttc cacatctcta ttttgacccg cagaataatc caaaatgcag atcggggatc
     8221 ccaccccacc caagaagaag cgcaaggtgg aggacgatcc cgtcgtttta caacgtcgtg
     8281 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     8341 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cggtcgactc
     8401 tagaggatcc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     8461 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     8521 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     8581 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     8641 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     8701 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     8761 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     8821 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     8881 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     8941 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     9001 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     9061 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     9121 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     9181 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     9241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     9301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     9361 tagttgtggt ttgtccaaac tcatcaatgt atcttaaagc ttatcgatac gcgtacggca
     9421 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     9481 agatcggccg gccttggcgc gcctagatct taatacgact cactataggg cgaattgggt
     9541 accg
//
