LOCUS       pBM_MCS_ALFA-str        7477 bp DNA     circular SYN 01-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 1, 2024 from SnapGene Viewer 7.1.1
            https://www.snapgene.com
COMMENT     This file is created by Vector NTI
            http://www.informaxinc.com/
FEATURES             Location/Qualifiers
     source          1..7477
                     /organism="synthetic DNA construct"
     source          5131..5223
     source          5224..5247
     rep_origin      2..457
                     /label=f1 origin
     promoter        484..588
                     /gene="bla"
                     /label=AmpR promoter
     promoter        490..588
                     /label=bla promoter
     CDS             589..1449
                     /codon_start=1
                     /label=Amp
                     /label=Amp(R)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1594..2267
                     /label=pUC origin
     rep_origin      1620..2208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_recomb     2511..2735
                     /label=Tn7R
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /label=Gm
                     /label=Gm(R)
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(3524..3551)
                     /label=Pc promoter
     misc_feature    3885..3905
                     /label=Link1
     promoter        3925..4052
                     /label=PH promoter
     promoter        3925..4016
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3941..3958
                     /label=Polyhedrin forward primer
     promoter        4189..4772
                     /label=CMV_immearly_promoter
     enhancer        4189..4568
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4569..4772
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        4712..4793
                     /label=CMV_promoter
     misc_feature    4908..5040
                     /label=chim_int
     intron          4908..5040
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             5080..5082
                     /codon_start=1
                     /label=Met
                     /translation="M"
     CDS             5083..5121
                     /codon_start=1
                     /label=ALFA
                     /translation="SRLEEELRRRLTE"
     CDS             5134..5157
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5167..5190
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5200..5223
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5224..5247
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5248..6168
                     /codon_start=1
                     /label=hUCP1
                     /translation="MGGLTASDVHPTLGVQLFSAGIAACLADVITFPLDTAKVRLQVQG
                     ECPTSSVIRYKGVLGTITAVVKTEGRMKLYSGLPAGLQRQISSASLRIGLYDTVQEFLT
                     AGKETAPSLGSKILAGLTTGGVAVFIGQPTEVVKVRLQAQSHLHGIKPRYTGTYNAYRI
                     IATTEGLTGLWKGTTPNLMRSVIINCTELVTYDLMKEAFVKNNILADDVPCHLVSALIA
                     GFCATAMSSPVDVVKTRFINSPPGQYKSVPNCAMKVFTNEGPTAFFKGLVPSFLRLGSW
                     NVIMFVCFEQLKRELSKSRQTMDCAT"
     CDS             6178..6195
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6202..6790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     misc_feature    6204..6779
                     /label=WPRE
     CDS             complement(6673..6684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6811..7051
                     /label=SV40 pA
     polyA_signal    6917..7051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    7058..7107
                     /label=Link2
     misc_recomb     7130..7295
                     /label=Tn7L
     mobile_element  7130..7295
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
      481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
      841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1981 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2281 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2341 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2401 cggtattttc tccttacgca tctgtgcggt atttcacacc gcagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtacttaat taacaacacc
     3901 atttgtactc cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa
     3961 ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc
     4021 cggattattc ataccgtccc accatcgggc gccaatattg gccattagcc atattattca
     4081 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     4141 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat
     4201 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     4261 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4321 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4381 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4441 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4501 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4561 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
     4621 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4681 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
     4741 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
     4801 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
     4861 cagtctcgaa cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg
     4921 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
     4981 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
     5041 gtgtccactc ccagttcaat tacagctctt aaggaattca tgtctcggct cgaggaagaa
     5101 ctgcgccgaa gactcactga gggtagtgga ggttggtctc atccacaatt tgagaaaggt
     5161 agtggagact acaaggacga tgatgacaag ggatcggggt ggagccaccc gcagttcgaa
     5221 aagctcgaag tgctgttcca aggccctatg gggggcctga cagcctcgga cgtacacccg
     5281 accctggggg tccagctctt ctcagctgga atagcggcgt gcttggcgga cgtgatcacc
     5341 ttcccgctgg acacggccaa agtccggctc caggtccaag gtgaatgccc gacgtccagt
     5401 gttattaggt ataaaggtgt cctgggaaca atcaccgctg tggtaaaaac agaagggcgg
     5461 atgaaactct acagcgggct gcctgcgggg cttcagcggc aaatcagctc cgcctctctc
     5521 aggatcggcc tctacgacac ggtccaggag ttcctcaccg cagggaaaga aacagcacct
     5581 agtttaggaa gcaagatttt agctggtcta acgactggag gagtggcagt attcattggg
     5641 caacccacag aggtcgtgaa agtcagactt caagcacaga gccatctcca cggaatcaaa
     5701 cctcgctaca cggggactta taatgcgtac agaataatag caacaaccga aggcttgacg
     5761 ggtctttgga aagggactac tcccaatctg atgagaagtg tcatcatcaa ttgtacagag
     5821 ctagtaacat atgatctaat gaaggaggcc tttgtgaaaa acaacatatt agcagatgac
     5881 gtcccctgcc acttggtgtc ggctcttatc gctggatttt gcgcaacagc tatgtcctcc
     5941 ccggtggatg tagtaaaaac cagatttatt aattctccac caggacagta caaaagtgtg
     6001 cccaactgtg caatgaaagt gttcactaac gaaggaccaa cggctttctt caaggggttg
     6061 gtaccttcct tcttgcgact tggatcctgg aacgtcatta tgtttgtgtg ctttgaacaa
     6121 ctgaaacgag aactgtcaaa gtcaaggcag actatggact gtgccacata actcgagcac
     6181 caccaccacc accactgaga taatcaacct ctggattaca aaatttgtga aagattgact
     6241 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     6301 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     6361 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     6421 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     6481 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     6541 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     6601 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     6661 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     6721 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     6781 gcctccccgc ctggtcgaga agtactagag gatcataatc agccatacca catttgtaga
     6841 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     6901 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6961 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7021 actcatcaat gtatcttatc atgtctggat ctgatcagta acaacaccat ttaaatggag
     7081 tggttacaaa tggagtggtt aattaagctg cttgagccta ggagatccga accagataag
     7141 tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac
     7201 acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca
     7261 cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg
     7321 tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctactttttc
     7381 tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg taattgactg
     7441 aatatcaacg cttatttgca gcctgaatgg cgaatgg
//
