LOCUS       Exported                6133 bp ds-DNA     circular SYN 17-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6133)
  AUTHORS   abeitz
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 21, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6133
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     109..128
                     /label=oSEQ054 pMXs-fwd-seq
     protein_bind    222..246
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     270..284
                     /label=oTA100 GG_entry_upstream_rev
     primer_bind     complement(270..284)
                     /label=oTA101 GG_entry_downstream_fwd
     primer_bind     308..336
                     /label=oEP168
     primer_bind     312..335
                     /label=oKL139
     misc_feature    312..320
                     /label=Kozak sequence
                     /note="Partial (missing last G) vertebrate consensus 
                     sequence for strong initiation of translation (Kozak, 
                     1987)"
     CDS             318..1064
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     318..337
                     /label=oCC09
     primer_bind     443..462
                     /label=oSEQ057 rTTa_fwd
     primer_bind     complement(495..508)
                     /label=oKL164
     primer_bind     complement(1020..1039)
                     /label=oSEQ058 rTTa_rev
     primer_bind     complement(1042..1061)
                     /label=oKL140
     primer_bind     complement(1042..1061)
                     /label=oKL140a
     primer_bind     complement(1042..1061)
                     /label=oCC13
     primer_bind     complement(1044..1068)
                     /label=oEP110
     primer_bind     1064..1092
                     /label=pPV3-none_fwd
     primer_bind     complement(1071..1104)
                     /label=pPV3-none_rev
     primer_bind     1103..1124
                     /label=oGE002 polyA_reporter_bb_fwd
     primer_bind     1105..1120
                     /label=oTA100 GG_entry_upstream_rev
     primer_bind     complement(1105..1120)
                     /label=oTA101 GG_entry_downstream_fwd
     protein_bind    complement(1114..1138)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1155..1743
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1321..1340)
                     /label=oSEQ034 WPRE_rev
     primer_bind     1578..1598
                     /label=oSEQ048 phage-dest-fwd-2
     CDS             complement(1626..1637)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1740..1757
                     /label=oSEQ013 flipped_WPRE_rev
     primer_bind     complement(1782..1800)
                     /label=oSEQ055 pMXs-rev-seq
     LTR             1923..2392
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     2589..2619
                     /label=pPV3-none_rev
     primer_bind     complement(2598..2622)
                     /label=pPV3-none_fwd
     rep_origin      complement(2693..3281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2773..2792)
                     /label=oSEQ067 pBR322-ori_fwd
     CDS             complement(3445..4305)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4068..4087
                     /label=oSEQ066 Amp_rev
     primer_bind     complement(4281..4305)
                     /label=oSEQ083 Amp-fwd
     promoter        complement(4306..4410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4352..4373
                     /label=oSEQ095 oKL157
     LTR             4440..5033
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5096..5453
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5512..5928
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5706..5725)
                     /label=oSEQ082 pMX-gag-rev
     primer_bind     5706..5725
                     /label=oSEQ096 pMX-gag_fwd
     misc_feature    join(5938..6133,1..179)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag
       61 gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt
      121 ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc
      181 cggatctagc tagttaatcg acggtatcga taagctatca aacaagtttg tacaaaaaag
      241 caggctccac catgggaacc aattcagtcg actggatccg gtactactga attcaagctt
      301 ggatcccaat ggccaccatg agccggctgg acaagagcaa agtgatcaac agcgccctgg
      361 aactgctgaa cggcgtgggc atcgagggcc tgaccacccg gaagctggcc cagaaactgg
      421 gcgtggaaca gcccaccctg tactggcacg tgaagaacaa gcgggccctg ctggacgccc
      481 tgcccatcga gatgctggac cggcaccaca cccacagctg ccctctggaa ggcgagagct
      541 ggcaggactt cctgcggaac aacgccaaga gctacagatg cgccctgctg agccaccggg
      601 acggcgccaa agtgcacctg ggcaccagac ccaccgagaa gcagtacgag acactggaaa
      661 accagctggc cttcctgtgc cagcagggct tcagcctgga aaacgccctg tacgccctga
      721 gcgccgtggg ccacttcacc ctgggctgcg tgctggaaga acaggaacac caggtcgcca
      781 aagaggaaag agagacaccc accaccgaca gcatgccccc cctgctgaag caggccatcg
      841 agctgttcga cagacagggc gccgagcccg ccttcctgtt cggcctggaa ctgatcatct
      901 gcggcctcga gaagcagctg aagtgcgaga gcggcggacc caccgacgcc ctggacgact
      961 tcgacctgga catgctgccc gccgatgccc tggatgattt tgatctggat atgctccctg
     1021 ctgacgctct cgatgacttc gatctcgata tgctgccagg ctaacaacgt tatcccctga
     1081 ttctgtggat aaccgtatta cgctgatatc tagacccagc tttcttgtac aaagtggttc
     1141 gatagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1201 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1261 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1321 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1381 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     1441 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1501 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     1561 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     1621 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     1681 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     1741 cgcatcgata ccgtcgacga taaaataaaa gattttattt agtctccaga aaaagggggg
     1801 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     1861 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     1921 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     1981 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     2041 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     2101 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     2161 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     2221 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     2281 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     2341 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     2401 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     2461 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2881 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3121 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3361 tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat
     3421 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3481 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3541 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3601 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3661 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3721 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3781 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3841 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3901 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3961 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4021 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4081 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4141 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4201 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4261 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4321 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4381 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcata
     4441 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     4501 tggaaaaata cataactgag aatagaaaag ttcagatcaa ggtcaggaac agatggaaca
     4561 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     4621 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     4681 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     4741 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4801 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     4861 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgattgactg agtcgcccgg
     4921 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     4981 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
     5041 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
     5101 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc
     5161 tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
     5221 agttcggaac acccggccgc aaccctggga gacgtcccag ggacttcggg ggccgttttt
     5281 gtggcccgac ctgagtccaa aaatcccgat cgttttggac tctttggtgc acccccctaa
     5341 taggagggat atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct
     5401 gaatttttgc tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc
     5461 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
     5521 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
     5581 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
     5641 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
     5701 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
     5761 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
     5821 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
     5881 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgcccc catatggcca
     5941 tatgagatct tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca
     6001 agagttacta acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac
     6061 gaagtctgga gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct
     6121 cacccttacc gag
//
