LOCUS       pTCB-nivoLC             5534 bp    DNA     circular SYN 03-APR-2025
DEFINITION  none.
ACCESSION   test
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5534)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 3, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1..5534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1..6
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct(1)
                     /note="/ApEinfo_fwdcolor=#00DF87"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_hidden"
                     /note="/ApEinfo_label=source:synthetic DNA construct"
                     /note="/ApEinfo_revcolor=#00DF87"
                     /note="/locus_tag=source:synthetic DNA construct(1)"
                     /organism="synthetic DNA construct"
     source          709..1547
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct(2)
                     /note="/ApEinfo_fwdcolor=#00DF87"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_hidden"
                     /note="/ApEinfo_label=source:synthetic DNA construct"
                     /note="/ApEinfo_revcolor=#00DF87"
                     /note="/locus_tag=source:synthetic DNA construct(2)"
                     /organism="synthetic DNA construct"
     source          1548..5534
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="/ApEinfo_fwdcolor=#00DF87"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_hidden"
                     /note="/ApEinfo_label=source:synthetic DNA construct"
                     /note="/ApEinfo_revcolor=#00DF87"
                     /note="/locus_tag=source:synthetic DNA construct"
                     /organism="synthetic DNA construct"
     misc_feature    1..6
                     /locus_tag="XhoI"
                     /label=XhoI
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=XhoI"
                     /note="/ApEinfo_revcolor=cyan"
     CDS             7..63
                     /codon_start=1
                     /label=leader sequence
                     /translation="MGWSCIILFLVATATGVHS"
     misc_feature    64..384
                     /label=Nivolumab VL
     CDS             385..708
                     /codon_start=1
                     /label=Ab CL
                     /translation="RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN
                     ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR
                     GEC"
     misc_feature    709..714
                     /locus_tag="NheI"
                     /label=NheI
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=NheI"
                     /note="/ApEinfo_revcolor=cyan"
     polyA_signal    758..879
                     /locus_tag="SV40 poly(A) signal"
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="/ApEinfo_fwdcolor=#ff3eee"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=SV40 poly(A) signal"
                     /note="/ApEinfo_revcolor=#ff3eee"
     terminator      994..1040
                     /locus_tag="T7 terminator(1)"
                     /label=T7 terminator(1)
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="/ApEinfo_fwdcolor=#9d1b1c"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=T7 terminator"
                     /note="/ApEinfo_revcolor=#9d1b1c"
     terminator      1070..1206
                     /locus_tag="rrnG terminator(1)"
                     /label=rrnG terminator(1)
                     /note="transcription terminator from the E. coli ribosomal
                     RNA rrnG operon (Albrechtsen et al., 1991)"
                     /note="/ApEinfo_fwdcolor=#9d1b1c"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=rrnG terminator"
                     /note="/ApEinfo_revcolor=#9d1b1c"
     misc_feature    1251..1322
                     /locus_tag="3' beta-globin insulator"
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
                     /note="/ApEinfo_fwdcolor=#7eff74"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=3' beta-globin insulator"
                     /note="/ApEinfo_revcolor=#7eff74"
     rep_origin      complement(1548..2136)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="/ApEinfo_fwdcolor=#999999"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=ori"
                     /note="/ApEinfo_revcolor=#999999"
     CDS             2882..3742
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /note="/ApEinfo_fwdcolor=#e9d024"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=AmpR"
                     /note="/ApEinfo_revcolor=#e9d024"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3755..3891
                     /locus_tag="rrnG terminator"
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal
                     RNA rrnG operon (Albrechtsen et al., 1991)"
                     /note="/ApEinfo_fwdcolor=#9d1b1c"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=rrnG terminator"
                     /note="/ApEinfo_revcolor=#9d1b1c"
     misc_feature    4043..4114
                     /locus_tag="5' beta-globin insulator"
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
                     /note="/ApEinfo_fwdcolor=#7eff74"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=5' beta-globin insulator"
                     /note="/ApEinfo_revcolor=#7eff74"
     terminator      4119..4165
                     /locus_tag="T7 terminator"
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="/ApEinfo_fwdcolor=#9d1b1c"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=T7 terminator"
                     /note="/ApEinfo_revcolor=#9d1b1c"
     polyA_signal    4183..4231
                     /locus_tag="synthetic polyadenylation signal"
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
                     /note="/ApEinfo_fwdcolor=#ff3eee"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=synthetic polyadenylation signal"
                     /note="/ApEinfo_revcolor=#ff3eee"
     enhancer        4312..4615
                     /locus_tag="CMV enhancer"
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="/ApEinfo_fwdcolor=#5ac3fa"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=CMV enhancer"
                     /note="/ApEinfo_revcolor=#5ac3fa"
     promoter        4617..4819
                     /locus_tag="CMV promoter"
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="/ApEinfo_fwdcolor=#346ee0"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=CMV promoter"
                     /note="/ApEinfo_revcolor=#346ee0"
     intron          4953..5428
                     /locus_tag="beta-globin intron"
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
                     /note="/ApEinfo_fwdcolor=#eb606c"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 0.5 0 1 2 0 0
                     -1 0
                     -0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
                     /note="/ApEinfo_label=beta-globin intron"
                     /note="/ApEinfo_revcolor=#eb606c"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tccgagatcg ttcttactca atctcctgca acgctttccc tcagtccagg agaacgcgcg
      121 accctttcat gcagagcctc tcagagtgtt tcctcatatc tcgcctggta tcagcaaaaa
      181 ccagggcaag caccccgctt gttgatttac gacgcgagca atcgggccac aggcattcca
      241 gccaggtttt caggaagcgg atcaggcacc gatttcacgc tgaccattag ctcccttgag
      301 ccagaagatt tcgcagtcta ctattgtcag caatcatcaa actggcctcg aaccttcggc
      361 cagggtacaa aagttgaaat aaagcgcacc gtcgccgcgc ccagcgtgtt catcttcccg
      421 cccagcgacg agcagctgaa gagcggcacg gccagcgtgg tctgcctgct caacaacttc
      481 tacccccggg aggccaaggt gcagtggaag gtggacaacg ccctgcagtc ggggaacagc
      541 caggagagcg tcaccgagca ggacagcaag gacagcacct acagcctgtc gagcaccctc
      601 acgctgagca aggccgacta cgagaagcac aaggtgtacg cgtgcgaggt gacccaccag
      661 ggcctgagca gccccgtcac caagtcgttc aaccgcggag aatgctgagc tagcttgact
      721 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
      781 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      841 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
      901 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
      961 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     1021 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     1081 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     1141 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     1201 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     1261 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     1321 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     1381 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     1441 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     1501 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1561 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1621 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1681 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1741 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1801 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1861 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1921 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1981 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2041 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2101 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2161 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2221 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2281 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2341 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     2401 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     2461 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     2521 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     2581 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     2641 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     2701 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     2761 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     2821 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     2881 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2941 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3001 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3061 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3121 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3181 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3241 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3301 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3361 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3421 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     3481 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3541 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3601 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3661 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3721 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     3781 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     3841 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     3901 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     3961 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     4021 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     4081 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     4141 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     4201 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     4261 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     4321 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     4381 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     4441 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     4501 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     4561 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     4621 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     4681 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     4741 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     4801 ggtctatata agcagagctg gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     4861 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctcccctcga agcttacatg
     4921 tggtaccgag ctcggatcct gagaacttca gggtgagtct atgggaccct tgatgttttc
     4981 tttccccttc ttttctatgg ttaagttcat gtcataggaa ggggagaagt aacagggtac
     5041 acatattgac caaatcaggg taattttgca tttgtaattt taaaaaatgc tttcttcttt
     5101 taatatactt ttttgtttat cttatttcta atactttccc taatctcttt ctttcagggc
     5161 aataatgata caatgtatca tgcctctttg caccattcta aagaataaca gtgataattt
     5221 ctgggttaag gcaatagcaa tatttctgca tataaatatt tctgcatata aattgtaact
     5281 gatgtaagag gtttcatatt gctaatagca gctacaatcc agctaccatt ctgcttttat
     5341 tttatggttg ggataaggct ggattattct gagtccaagc taggcccttt tgctaatcat
     5401 gttcatacct cttatcttcc tcccacagct cctgggcaac gtgctggtct gtgtgctggc
     5461 ccatcacttt ggcaaaggcg gccgctgcca agcttccgag ctctcgaatt caaaggaggt
     5521 accgcgatat ctac
//
