LOCUS       pMXs-9SA.mNgn2-x        6333 bp DNA     circular SYN 25-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2025 from SnapGene 7.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     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"
     misc_feature    260..280
                     /label=pENTR-Gibson-REV
     misc_feature    289..306
                     /label=MCS
     misc_feature    311..316
                     /label=Kozak sequence
                     /note="Partial (missing last G) vertebrate consensus
                     sequence for strong initiation of translation (Kozak, 
                     1987)"
     CDS             317..1105
                     /codon_start=1
                     /label=phospho-mut-mNgn2
                     /translation="MFVKSETLELKEEEEVLMLLGSAAPASATLTPMSSSADEEEDEEL
                     RRPGSARGQRGAEAGQGVQGAPASGAGGCRPGRLLGLMHECKRRPSRSRAVSRGAKTAE
                     TVQRIKKTRRLKANNRERNRMHNLNAALDALREVLPTFPEDAKLTKIETLRFAHNYIWA
                     LTETLRLADHCAGAGGLQGALFTEAVLLAPGAALGASGDAPAPPSSWSCTNAPASSSNS
                     TAPYSCTLAPAAPGSDVDYWQPPPPEKHRYAPHLPLARDCI"
     misc_feature    386..391
                     /label=SP -> AP
     misc_feature    512..517
                     /label=SP -> AP
     misc_feature    890..895
                     /label=SP -> AP
     misc_feature    923..928
                     /label=SP -> AP
     misc_feature    929..934
                     /label=SP -> AP
     misc_feature    959..964
                     /label=SP -> AP
     misc_feature    986..991
                     /label=SP -> AP
     misc_feature    1007..1012
                     /label=SP -> AP
     misc_feature    1016..1021
                     /label=SP -> AP
     CDS             1145..1243
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=3xHA
     protein_bind    complement(1314..1338)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1355..1943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2123..2592
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(2893..3481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3645..4505)
                     /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"
     promoter        complement(4506..4610)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4640..5233
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    5296..5653
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             5712..6128
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    join(6138..6333,1..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 ggatccaatg gccaccatgt tcgtcaaatc tgagactctg gagttgaagg aggaagagga
      361 ggtactgatg ctgctgggct cggctgcccc ggcctcggcg accctgaccc cgatgtcctc
      421 cagcgcggac gaggaggagg acgaggagct gcgccggccg ggctccgcgc gtgggcagcg
      481 tggagcggaa gccgggcagg gggtgcaggg cgccccggcg tcgggtgccg ggggttgccg
      541 gccagggcgg ctgctgggcc tgatgcacga gtgcaagcgt cgcccgtcgc gctcacgggc
      601 cgtctcccga ggtgccaaga cggcggagac ggtgcagcgc atcaagaaga cccgcaggct
      661 caaggccaac aaccgcgagc gcaaccgcat gcacaaccta aacgccgcgc tggacgcgct
      721 gcgcgaggtg ctgcccacct tccccgagga tgccaagctc acgaagatcg agacgctgcg
      781 cttcgcccac aattacatct gggcgctcac cgagactctg cgcctggcgg accactgcgc
      841 cggcgccggt ggcctccagg gggcgctctt cacggaggcg gtgctcctgg ccccgggagc
      901 tgcgctcggc gccagcgggg acgcccctgc cccaccttcc tcctggagct gcaccaacgc
      961 cccggcgtca tcctccaact ccacggcccc atacagctgc actttagccc ccgctgcccc
     1021 cgggtcagac gtggactact ggcagccccc acctccggag aagcatcgtt atgcgcctca
     1081 cctgcccctc gccagggact gtatcgggtc tggctccgga tcaagtggtg gcagcggatc
     1141 aggatatcca tacgatgtac cagactatgc gggatcgggt tacccctacg acgtccccga
     1201 ctacgccggc tccggctacc cctacgacgt ccctgattat gctggttctg gtggtgttga
     1261 ataacaacgt tatcccctga ttctgtggat aaccgtatta cgctgatatc tagacccagc
     1321 tttcttgtac aaagtggttc gatagcttat cgataatcaa cctctggatt acaaaatttg
     1381 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1441 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1501 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1561 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1621 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1681 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     1741 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     1801 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     1861 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     1921 ctccctttgg gccgcctccc cgcatcgata ccgtcgacga taaaataaaa gattttattt
     1981 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     2041 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca
     2101 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     2161 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
     2221 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     2281 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     2341 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     2401 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct
     2461 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     2521 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     2581 gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     2641 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2701 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2761 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2821 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2881 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2941 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3001 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3061 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3121 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3181 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3241 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3301 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3361 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3421 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3481 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc
     3601 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3661 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3721 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3781 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3841 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3901 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3961 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4021 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4081 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4141 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4201 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4261 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4321 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4381 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4441 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4501 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4561 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4621 cgaaaagtgc cacctgcata atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4681 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagaaaag ttcagatcaa
     4741 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     4801 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     4861 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     4921 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     4981 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     5041 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     5101 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     5161 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     5221 ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac
     5281 ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc
     5341 tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg
     5401 gacccgtggt ggaactgacg agttcggaac acccggccgc aaccctggga gacgtcccag
     5461 ggacttcggg ggccgttttt gtggcccgac ctgagtccaa aaatcccgat cgttttggac
     5521 tctttggtgc acccccctaa taggagggat atgtggttct ggtaggagac gagaacctaa
     5581 aacagttccc gcctccgtct gaatttttgc tttcggtttg ggaccgaagc cgcgccgcgc
     5641 gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     5701 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     5761 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     5821 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     5881 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     5941 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     6001 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     6061 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     6121 taggcgcccc catatggcca tatgagatct tatatggggc acccccgccc cttgtaaact
     6181 tccctgaccc tgacatgaca agagttacta acagcccctc tctccaagct cacttacagg
     6241 ctctctactt agtccagcac gaagtctgga gacctctggc ggcagcctac caagaacaac
     6301 tggaccgacc ggtggtacct cacccttacc gag
//
